SEC Info℠  
    Home      Search      My Interests      Help      Sign In      Please Sign In

Trupanion, Inc. – ‘10-K’ for 12/31/20 – ‘EX-101.DEF’

On:  Friday, 2/12/21, at 8:22am ET   ·   For:  12/31/20   ·   Accession #:  1371285-21-41   ·   File #:  1-36537

Previous ‘10-K’:  ‘10-K’ on 2/14/20 for 12/31/19   ·   Next:  ‘10-K’ on 2/17/22 for 12/31/21   ·   Latest:  ‘10-K’ on 2/27/24 for 12/31/23   ·   21 References:   

Find Words in Filings emoji
 		  in    Show  and   ↓Hints

  As Of               Filer                 Filing    For·On·As Docs:Size

 2/12/21  Trupanion, Inc.                   10-K       12/31/20  101:12M

Annual Report   —   Form 10-K
Filing Table of Contents

Document/Exhibit                   Description                      Pages   Size 

 1: 10-K        Annual Report                                       HTML   1.51M 
 2: EX-3.3      Articles of Incorporation/Organization or Bylaws    HTML    113K 
 3: EX-10.16    Material Contract                                   HTML     31K 
 4: EX-10.17    Material Contract                                   HTML     41K 
 5: EX-21.1     Subsidiaries List                                   HTML     28K 
 6: EX-23.1     Consent of Expert or Counsel                        HTML     27K 
 7: EX-31.1     Certification -- §302 - SOA'02                      HTML     29K 
 8: EX-31.2     Certification -- §302 - SOA'02                      HTML     29K 
 9: EX-32.1     Certification -- §906 - SOA'02                      HTML     26K 
10: EX-32.2     Certification -- §906 - SOA'02                      HTML     26K 
17: R1          Cover Page Document                                 HTML     87K 
18: R2          Consolidated Statement of Operations                HTML     79K 
19: R3          Consolidated Statement of Comprehensive Income      HTML     43K 
                Statement                                                        
20: R4          Consolidated Balance Sheet                          HTML    111K 
21: R5          Consolidated Balance Sheet Condensed Consolidated   HTML     49K 
                Balance Sheet Parentheticals                                     
22: R6          Consolidated Statement of Stockholders' Equity      HTML     86K 
                Statement                                                        
23: R7          Consolidated Statement of Cash Flows                HTML    117K 
24: R8          Condensed Consolidated Balance Sheet                HTML     54K 
                Parentheticals                                                   
25: R9          Schedule 1-Parent Only Disclosures [Schedule]       HTML     77K 
                Condensed Statement of Comprehensive Loss                        
                (Details)                                                        
26: R10         Net Loss per Share                                  HTML     35K 
27: R11         Business Combinations                               HTML     36K 
28: R12         Property Plant and Equipment (Notes)                HTML     40K 
29: R13         Intangible Assets (Notes)                           HTML     57K 
30: R14         Investment Securities (Notes)                       HTML     73K 
31: R15         Other Investments (Notes)                           HTML     30K 
32: R16         Fair Value                                          HTML     65K 
33: R17         Commitment and Contingencies                        HTML     46K 
34: R18         Claims Reserve (Notes)                              HTML    172K 
35: R19         Debt                                                HTML     28K 
36: R20         Stock-based Compensation                            HTML     89K 
37: R21         Leases (Notes)                                      HTML     36K 
38: R22         Stockholder's Equity (Notes)                        HTML     31K 
39: R23         Segments                                            HTML     71K 
40: R24         Dividend Restrictions Statutory Surplus (Notes)     HTML     38K 
41: R25         Income Taxes (Notes)                                HTML    102K 
42: R26         Retirement Plan (Notes)                             HTML     28K 
43: R27         Related Party Disclosures                           HTML     29K 
44: R28         Quarterly Financial Information (Notes)             HTML     62K 
45: R29         Schedule 1-Parent Only Disclosures [Schedule]       HTML    134K 
                (Notes)                                                          
46: R30         Nature of Operations and Summary of Significant     HTML    165K 
                Accounting Policies (Policies)                                   
47: R31         Other Investments details (Policies)                HTML     32K 
48: R32         Net Loss per Share (Tables)                         HTML     34K 
49: R33         Property Plant and Equipment (Tables)               HTML     41K 
50: R34         Investment Securities Available-for-Sale (Tables)   HTML     75K 
51: R35         Fair Value (Tables)                                 HTML     61K 
52: R36         Commitment and Contingencies (Tables)               HTML     44K 
53: R37         Claims Reserve (Tables)                             HTML    134K 
54: R38         Stock-based Compensation (Tables)                   HTML     98K 
55: R39         Segments (Tables)                                   HTML     68K 
56: R40         Dividend Restrictions Statutory Surplus (Tables)    HTML     36K 
57: R41         Income Taxes (Tables)                               HTML    105K 
58: R42         Quarterly Financial Information (Tables)            HTML     62K 
59: R43         Nature of Operations and Summary of Significant     HTML     69K 
                Accounting Policies Narrative (Details)                          
60: R44         Nature of Operations and Summary of Significant     HTML     32K 
                Accounting Policies Deferred Acquisition Costs                   
                (Details)                                                        
61: R45         Nature of Operations and Summary of Significant     HTML     39K 
                Accounting Policies Property, Plant, and                         
                Equipment, Useful Life (Details)                                 
62: R46         Net Loss per Share (Details) Schedule of            HTML     39K 
                Antidilutive Securities Excluded from Computation                
                of Earnings Per Share                                            
63: R47         Business Combinations (Details)                     HTML     53K 
64: R48         Property Plant and Equipment (Details)              HTML     48K 
65: R49         Property Plant and Equipment Real Estate            HTML     34K 
                Acquisition (Details)                                            
66: R50         Intangible Assets (Details)                         HTML     94K 
67: R51         Investment Securities (Details) Investment          HTML     60K 
                Schedule                                                         
68: R52         Investment Securities (Details) Available-for-Sale  HTML     36K 
69: R53         Other Investments (Details)                         HTML     36K 
70: R54         Fair Value (Details) Unobservable                   HTML     79K 
71: R55         Commitment and Contingencies Commitments and        HTML     69K 
                Contingencies (Details)                                          
72: R56         Claims Reserve (Details)                            HTML     66K 
73: R57         Claims Reserve Cumulative claims paid and claims    HTML     62K 
                adjustment expenses (Details)                                    
74: R58         Claims Reserve Incurred claims and claim            HTML     53K 
                adjustment expense (Details)                                     
75: R59         Debt (Details) Narrative                            HTML     31K 
76: R60         Stock-based Compensation (Details) Narrative        HTML     44K 
77: R61         Stock-based Compensation Expense Category           HTML     55K 
                (Details)                                                        
78: R62         Stock-based Compensation Valuation Assumptions      HTML     41K 
                (Details)                                                        
79: R63         Stock-based Compensation Option Activity (Details)  HTML     68K 
80: R64         Stock-based Compensation Options Granted (Details)  HTML     30K 
81: R65         Stock-based Compensation Restricted Stock Awards    HTML     51K 
                (Details)                                                        
82: R66         Leases (Details)                                    HTML     47K 
83: R67         Stockholder's Equity Narrative (Details)            HTML     40K 
84: R68         Segments (Details) Business Segment                 HTML     68K 
85: R69         Segments (Details) Revenue by Geography             HTML     38K 
86: R70         Dividend Restrictions Statutory Surplus (Details)   HTML     39K 
87: R71         Income Taxes Income before taxes (Details)          HTML     34K 
88: R72         Income Taxes Income tax benefits (Details)          HTML     46K 
89: R73         Income Taxes Tax Rate Reconciliation (Details)      HTML     49K 
90: R74         Income Taxes Deferred tax assets and liabilities    HTML     96K 
                (Details)                                                        
91: R75         Income Taxes Unrecognized Tax Benefits (Details)    HTML     43K 
92: R76         Retirement Plan Details (Details)                   HTML     27K 
93: R77         Related Party Disclosures (Details)                 HTML     32K 
94: R78         Quarterly Financial Information (Details)           HTML     71K 
95: R79         Schedule 1-Parent Only Disclosures [Schedule]       HTML    111K 
                Condensed Balance Sheet (Details)                                
96: R80         Schedule 1-Parent Only Disclosures [Schedule]       HTML     98K 
                Condensed Statements of Cash Flows (Details)                     
97: R9999       Uncategorized Items - trup-20201231.htm             HTML     27K 
99: XML         IDEA XML File -- Filing Summary                      XML    177K 
16: XML         XBRL Instance -- trup-20201231_htm                   XML   2.79M 
98: EXCEL       IDEA Workbook of Financial Reports                  XLSX    120K 
12: EX-101.CAL  XBRL Calculations -- trup-20201231_cal               XML    288K 
13: EX-101.DEF  XBRL Definitions -- trup-20201231_def                XML    650K 
14: EX-101.LAB  XBRL Labels -- trup-20201231_lab                     XML   1.88M 
15: EX-101.PRE  XBRL Presentations -- trup-20201231_pre              XML   1.20M 
11: EX-101.SCH  XBRL Schema -- trup-20201231                         XSD    187K 
100: JSON        XBRL Instance as JSON Data -- MetaLinks              435±   665K  
101: ZIP         XBRL Zipped Folder -- 0001371285-21-000041-xbrl      Zip    640K

‘EX-101.DEF’   —   XBRL Definitions — trup-20201231_def

This Exhibit is an XBRL XML File.

                                                                                                                                                                                
<?xml version="1.0" encoding="windows-1252"?>
<!-- XBRL Document Created with Wdesk from Workiva -->
<!-- Copyright 2021 Workiva -->
<!-- r:83e64ae9-9748-42df-883b-d8ee35b3cee3,g:39b4dd84-186b-4da6-b716-294840996b2a -->
<link:linkbase xmlns:link="http://www.xbrl.org/2003/linkbase" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xmlns:xbrldt="http://xbrl.org/2005/xbrldt" xsi:schemaLocation="http://www.xbrl.org/2003/linkbase http://www.xbrl.org/2003/xbrl-linkbase-2003-12-31.xsd">
<link:arcroleRef arcroleURI="http://xbrl.org/int/dim/arcrole/all" xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#all"/>
<link:arcroleRef arcroleURI="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#hypercube-dimension"/>
<link:arcroleRef arcroleURI="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#dimension-default"/>
<link:arcroleRef arcroleURI="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#dimension-domain"/>
<link:arcroleRef arcroleURI="http://xbrl.org/int/dim/arcrole/domain-member" xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#domain-member"/>
<link:roleRef roleURI="http://www.trupanion.com/role/CoverPageDocument" xlink:type="simple" xlink:href="trup-20201231.xsd#CoverPageDocument"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/CoverPageDocument" xlink:type="extended" id="i259de9e39cbf49aaa6d8d074af7991d1_CoverPageDocument"/>
<link:roleRef roleURI="http://www.trupanion.com/role/ConsolidatedStatementofOperations" xlink:type="simple" xlink:href="trup-20201231.xsd#ConsolidatedStatementofOperations"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ConsolidatedStatementofOperations" xlink:type="extended" id="i02460ff084ff47318bceae6e268b3b93_ConsolidatedStatementofOperations"/>
<link:roleRef roleURI="http://www.trupanion.com/role/ConsolidatedStatementofComprehensiveIncomeStatement" xlink:type="simple" xlink:href="trup-20201231.xsd#ConsolidatedStatementofComprehensiveIncomeStatement"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ConsolidatedStatementofComprehensiveIncomeStatement" xlink:type="extended" id="i4192ad6788c441ce99936e534c1bdb3b_ConsolidatedStatementofComprehensiveIncomeStatement">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementLineItems_ab0ba737-e2ce-42a8-89b0-8d59a2b4f18f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementTable_0ebd63b8-fdca-49d8-af3b-0c76086e95a7" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_StatementLineItems_ab0ba737-e2ce-42a8-89b0-8d59a2b4f18f" xlink:to="loc_us-gaap_StatementTable_0ebd63b8-fdca-49d8-af3b-0c76086e95a7" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_StatementScenarioAxis_ab6d19a1-2949-41bc-a0b1-443e3ad98f16" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_StatementScenarioAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_StatementTable_0ebd63b8-fdca-49d8-af3b-0c76086e95a7" xlink:to="loc_srt_StatementScenarioAxis_ab6d19a1-2949-41bc-a0b1-443e3ad98f16" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ScenarioUnspecifiedDomain_ab6d19a1-2949-41bc-a0b1-443e3ad98f16_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ScenarioUnspecifiedDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_StatementScenarioAxis_ab6d19a1-2949-41bc-a0b1-443e3ad98f16" xlink:to="loc_srt_ScenarioUnspecifiedDomain_ab6d19a1-2949-41bc-a0b1-443e3ad98f16_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ScenarioUnspecifiedDomain_48d2f8d5-e759-48c7-b3be-c0d5cd62c429" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ScenarioUnspecifiedDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_StatementScenarioAxis_ab6d19a1-2949-41bc-a0b1-443e3ad98f16" xlink:to="loc_srt_ScenarioUnspecifiedDomain_48d2f8d5-e759-48c7-b3be-c0d5cd62c429" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/ConsolidatedBalanceSheet" xlink:type="simple" xlink:href="trup-20201231.xsd#ConsolidatedBalanceSheet"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ConsolidatedBalanceSheet" xlink:type="extended" id="i56e924969394413db93ebe20c041348c_ConsolidatedBalanceSheet">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementLineItems_7af6a4ca-a644-42c4-85bc-f03a365427e3" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementTable_49f0f854-8dd0-42fe-93ed-d3d2750f076f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_StatementLineItems_7af6a4ca-a644-42c4-85bc-f03a365427e3" xlink:to="loc_us-gaap_StatementTable_49f0f854-8dd0-42fe-93ed-d3d2750f076f" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_StatementScenarioAxis_afab9dde-be47-4e05-bb3e-73289103fe07" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_StatementScenarioAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_StatementTable_49f0f854-8dd0-42fe-93ed-d3d2750f076f" xlink:to="loc_srt_StatementScenarioAxis_afab9dde-be47-4e05-bb3e-73289103fe07" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ScenarioUnspecifiedDomain_afab9dde-be47-4e05-bb3e-73289103fe07_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ScenarioUnspecifiedDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_StatementScenarioAxis_afab9dde-be47-4e05-bb3e-73289103fe07" xlink:to="loc_srt_ScenarioUnspecifiedDomain_afab9dde-be47-4e05-bb3e-73289103fe07_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ScenarioUnspecifiedDomain_ee8b79c0-bbf3-43b1-9ea0-326cb5bce6a7" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ScenarioUnspecifiedDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_StatementScenarioAxis_afab9dde-be47-4e05-bb3e-73289103fe07" xlink:to="loc_srt_ScenarioUnspecifiedDomain_ee8b79c0-bbf3-43b1-9ea0-326cb5bce6a7" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementClassOfStockAxis_9ff77e57-f688-44ce-a44f-b3d0e0b1acda" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementClassOfStockAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_StatementTable_49f0f854-8dd0-42fe-93ed-d3d2750f076f" xlink:to="loc_us-gaap_StatementClassOfStockAxis_9ff77e57-f688-44ce-a44f-b3d0e0b1acda" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_9ff77e57-f688-44ce-a44f-b3d0e0b1acda_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementClassOfStockAxis_9ff77e57-f688-44ce-a44f-b3d0e0b1acda" xlink:to="loc_us-gaap_ClassOfStockDomain_9ff77e57-f688-44ce-a44f-b3d0e0b1acda_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_e9afbb0e-df73-4cf3-896e-47c62ef80a8a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementClassOfStockAxis_9ff77e57-f688-44ce-a44f-b3d0e0b1acda" xlink:to="loc_us-gaap_ClassOfStockDomain_e9afbb0e-df73-4cf3-896e-47c62ef80a8a" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/ConsolidatedBalanceSheetCondensedConsolidatedBalanceSheetParentheticals" xlink:type="simple" xlink:href="trup-20201231.xsd#ConsolidatedBalanceSheetCondensedConsolidatedBalanceSheetParentheticals"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ConsolidatedBalanceSheetCondensedConsolidatedBalanceSheetParentheticals" xlink:type="extended" id="i84c239e19d2640e394f99c17432327bc_ConsolidatedBalanceSheetCondensedConsolidatedBalanceSheetParentheticals">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockParOrStatedValuePerShare_a9f47454-5149-49ec-9248-ab65b91c5ab9" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockParOrStatedValuePerShare"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_CommonStockParOrStatedValuePerShare_a9f47454-5149-49ec-9248-ab65b91c5ab9" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockSharesAuthorized_9d2ac487-9dd1-4031-86b4-03c360a8f624" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockSharesAuthorized"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_CommonStockSharesAuthorized_9d2ac487-9dd1-4031-86b4-03c360a8f624" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockSharesIssued_14b37ae1-564e-4f0c-8e09-e24f0f7a0a9a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockSharesIssued"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_CommonStockSharesIssued_14b37ae1-564e-4f0c-8e09-e24f0f7a0a9a" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockSharesOutstanding_852dc8db-0200-4017-8158-24308711814c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockSharesOutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_CommonStockSharesOutstanding_852dc8db-0200-4017-8158-24308711814c" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockParOrStatedValuePerShare_325875ce-8a3b-4e43-873e-96777a7cb395" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockParOrStatedValuePerShare"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_PreferredStockParOrStatedValuePerShare_325875ce-8a3b-4e43-873e-96777a7cb395" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockSharesAuthorized_fe89fa43-323d-4e09-924f-e4bd2ad93178" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockSharesAuthorized"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_PreferredStockSharesAuthorized_fe89fa43-323d-4e09-924f-e4bd2ad93178" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockSharesIssued_6b1b5ba9-3703-47e0-bf12-c98e317f7ced" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockSharesIssued"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_PreferredStockSharesIssued_6b1b5ba9-3703-47e0-bf12-c98e317f7ced" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockSharesOutstanding_285be00d-1860-491f-8c9c-76ba27cee680" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockSharesOutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_PreferredStockSharesOutstanding_285be00d-1860-491f-8c9c-76ba27cee680" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TreasuryStockShares_cb967229-77fe-4735-8b46-8ecadf00e8a1" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TreasuryStockShares"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_TreasuryStockShares_cb967229-77fe-4735-8b46-8ecadf00e8a1" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementTable_9ca5e3cb-7003-423c-8248-90369a85d485" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_StatementLineItems_ab1320df-f948-430a-acf9-441b0b05ff1f" xlink:to="loc_us-gaap_StatementTable_9ca5e3cb-7003-423c-8248-90369a85d485" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementClassOfStockAxis_b085e2aa-39c2-451e-a7fe-4f3d6a6ca1e6" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementClassOfStockAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_StatementTable_9ca5e3cb-7003-423c-8248-90369a85d485" xlink:to="loc_us-gaap_StatementClassOfStockAxis_b085e2aa-39c2-451e-a7fe-4f3d6a6ca1e6" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_b085e2aa-39c2-451e-a7fe-4f3d6a6ca1e6_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementClassOfStockAxis_b085e2aa-39c2-451e-a7fe-4f3d6a6ca1e6" xlink:to="loc_us-gaap_ClassOfStockDomain_b085e2aa-39c2-451e-a7fe-4f3d6a6ca1e6_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_f516686d-9260-45f9-90a3-39a5a71bbbdb" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementClassOfStockAxis_b085e2aa-39c2-451e-a7fe-4f3d6a6ca1e6" xlink:to="loc_us-gaap_ClassOfStockDomain_f516686d-9260-45f9-90a3-39a5a71bbbdb" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockMember_8ebc8410-8d15-47a4-891e-1456639d5745" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_f516686d-9260-45f9-90a3-39a5a71bbbdb" xlink:to="loc_us-gaap_CommonStockMember_8ebc8410-8d15-47a4-891e-1456639d5745" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockMember_ddc558ea-a4e6-4787-a78c-25f0bcb3bc93" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_f516686d-9260-45f9-90a3-39a5a71bbbdb" xlink:to="loc_us-gaap_PreferredStockMember_ddc558ea-a4e6-4787-a78c-25f0bcb3bc93" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TreasuryStockMember_3e1a9343-0b96-4329-8509-89d9416f9613" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TreasuryStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_f516686d-9260-45f9-90a3-39a5a71bbbdb" xlink:to="loc_us-gaap_TreasuryStockMember_3e1a9343-0b96-4329-8509-89d9416f9613" xlink:type="arc" order="2"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/ConsolidatedStatementofStockholdersEquityStatement" xlink:type="simple" xlink:href="trup-20201231.xsd#ConsolidatedStatementofStockholdersEquityStatement"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ConsolidatedStatementofStockholdersEquityStatement" xlink:type="extended" id="i0777ccdb90024e8da77e6be123f5e77b_ConsolidatedStatementofStockholdersEquityStatement">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StockIssuedDuringPeriodSharesNewIssues_9eeb55eb-205a-4584-8d5a-391e14578137" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StockIssuedDuringPeriodSharesNewIssues"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:to="loc_us-gaap_StockIssuedDuringPeriodSharesNewIssues_9eeb55eb-205a-4584-8d5a-391e14578137" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Proceedsfromissuanceofcommonstockfollowonpublicoffering_f69105ea-f68c-41a2-818f-dec912b6af5f" xlink:href="trup-20201231.xsd#trup_Proceedsfromissuanceofcommonstockfollowonpublicoffering"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:to="loc_trup_Proceedsfromissuanceofcommonstockfollowonpublicoffering_f69105ea-f68c-41a2-818f-dec912b6af5f" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_PaymenttoAcquireCorporateBuildingShares_f30376e8-a6a3-437b-a612-ed15d659e76f" xlink:href="trup-20201231.xsd#trup_PaymenttoAcquireCorporateBuildingShares"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:to="loc_trup_PaymenttoAcquireCorporateBuildingShares_f30376e8-a6a3-437b-a612-ed15d659e76f" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_PaymenttoAcquireCorporateBuildingFairValueofShares_a5b0ed9a-581c-4cbb-9675-c1bdddf9fff5" xlink:href="trup-20201231.xsd#trup_PaymenttoAcquireCorporateBuildingFairValueofShares"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:to="loc_trup_PaymenttoAcquireCorporateBuildingFairValueofShares_a5b0ed9a-581c-4cbb-9675-c1bdddf9fff5" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_StockIssuedDuringPeriodSharesSharebasedCompensationNetofTaxWithholdings_682d3b30-1d06-4d94-b4d0-91c914bab59e" xlink:href="trup-20201231.xsd#trup_StockIssuedDuringPeriodSharesSharebasedCompensationNetofTaxWithholdings"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:to="loc_trup_StockIssuedDuringPeriodSharesSharebasedCompensationNetofTaxWithholdings_682d3b30-1d06-4d94-b4d0-91c914bab59e" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_StockIssuedDuringPeriodValueSharebasedCompensationNetofTaxWithholdings_f61291c1-4efb-47d2-ba88-36c013a65e19" xlink:href="trup-20201231.xsd#trup_StockIssuedDuringPeriodValueSharebasedCompensationNetofTaxWithholdings"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:to="loc_trup_StockIssuedDuringPeriodValueSharebasedCompensationNetofTaxWithholdings_f61291c1-4efb-47d2-ba88-36c013a65e19" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TreasuryStockSharesAcquired_3de2d609-6ebc-4f01-8f57-0219dc0102ac" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TreasuryStockSharesAcquired"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:to="loc_us-gaap_TreasuryStockSharesAcquired_3de2d609-6ebc-4f01-8f57-0219dc0102ac" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncreaseDecreaseInStockholdersEquityRollForward"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:to="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StockholdersEquity_18906eb9-fb4f-4e45-9c03-c51857f1d3f1" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StockholdersEquity"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_us-gaap_StockholdersEquity_18906eb9-fb4f-4e45-9c03-c51857f1d3f1" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockSharesOutstanding_cc4433bc-5ad4-452d-b7be-399d53a5f4d6" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockSharesOutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_us-gaap_CommonStockSharesOutstanding_cc4433bc-5ad4-452d-b7be-399d53a5f4d6" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Redemptionofwarrants_2850896d-f382-4249-a618-b83fd0fb430b" xlink:href="trup-20201231.xsd#trup_Redemptionofwarrants"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_trup_Redemptionofwarrants_2850896d-f382-4249-a618-b83fd0fb430b" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Settlementofwarrantliabilities_6a349b72-9e61-41ea-8aa6-5f78920fadce" xlink:href="trup-20201231.xsd#trup_Settlementofwarrantliabilities"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_trup_Settlementofwarrantliabilities_6a349b72-9e61-41ea-8aa6-5f78920fadce" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StockIssuedDuringPeriodValueNewIssues_11f5c868-7be2-4d33-b203-87b02a8cb1c2" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StockIssuedDuringPeriodValueNewIssues"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_us-gaap_StockIssuedDuringPeriodValueNewIssues_11f5c868-7be2-4d33-b203-87b02a8cb1c2" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AdjustmentsToAdditionalPaidInCapitalSharebasedCompensationRequisiteServicePeriodRecognitionValue_e5f63111-ad89-4d8c-a884-e562ef2a48ed" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AdjustmentsToAdditionalPaidInCapitalSharebasedCompensationRequisiteServicePeriodRecognitionValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_us-gaap_AdjustmentsToAdditionalPaidInCapitalSharebasedCompensationRequisiteServicePeriodRecognitionValue_e5f63111-ad89-4d8c-a884-e562ef2a48ed" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TreasuryStockValueAcquiredCostMethod_34acbdd0-696e-40f1-a705-39d459a582c7" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TreasuryStockValueAcquiredCostMethod"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_us-gaap_TreasuryStockValueAcquiredCostMethod_34acbdd0-696e-40f1-a705-39d459a582c7" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent_2341e00a-ccde-4505-9566-889f254e714e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent_2341e00a-ccde-4505-9566-889f254e714e" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTax_e56c0efd-9bc4-4297-8b7e-0a1dbcdff6ce" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherComprehensiveIncomeLossNetOfTax"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTax_e56c0efd-9bc4-4297-8b7e-0a1dbcdff6ce" xlink:type="arc" order="9"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetIncomeLossAvailableToCommonStockholdersBasic_5e03be53-6770-402c-9112-c47b0214454b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_NetIncomeLossAvailableToCommonStockholdersBasic"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncreaseDecreaseInStockholdersEquityRollForward_e0c38416-5938-40a9-8835-39635dc07b02" xlink:to="loc_us-gaap_NetIncomeLossAvailableToCommonStockholdersBasic_5e03be53-6770-402c-9112-c47b0214454b" xlink:type="arc" order="10"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StockholdersEquity_fd19f8b9-bc3e-477f-83c8-5d436b5c9b2f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StockholdersEquity"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementTable_31be8916-df85-47a3-95a3-fdee175fa97c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_StatementLineItems_4a90396d-9c0f-43ee-aae9-0d45c7993711" xlink:to="loc_us-gaap_StatementTable_31be8916-df85-47a3-95a3-fdee175fa97c" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementEquityComponentsAxis_a9b56698-d33b-4482-99d8-fc06b13da3cc" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementEquityComponentsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_StatementTable_31be8916-df85-47a3-95a3-fdee175fa97c" xlink:to="loc_us-gaap_StatementEquityComponentsAxis_a9b56698-d33b-4482-99d8-fc06b13da3cc" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_EquityComponentDomain_a9b56698-d33b-4482-99d8-fc06b13da3cc_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_EquityComponentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementEquityComponentsAxis_a9b56698-d33b-4482-99d8-fc06b13da3cc" xlink:to="loc_us-gaap_EquityComponentDomain_a9b56698-d33b-4482-99d8-fc06b13da3cc_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_EquityComponentDomain_bc771a3a-8eb0-4205-acbc-c6f477c9e9ac" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_EquityComponentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementEquityComponentsAxis_a9b56698-d33b-4482-99d8-fc06b13da3cc" xlink:to="loc_us-gaap_EquityComponentDomain_bc771a3a-8eb0-4205-acbc-c6f477c9e9ac" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockMember_569bc24b-119f-4a2e-8179-38dd91010f28" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EquityComponentDomain_bc771a3a-8eb0-4205-acbc-c6f477c9e9ac" xlink:to="loc_us-gaap_PreferredStockMember_569bc24b-119f-4a2e-8179-38dd91010f28" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockMember_8ab80ab8-f2b6-4e25-8c20-5f38670a6386" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EquityComponentDomain_bc771a3a-8eb0-4205-acbc-c6f477c9e9ac" xlink:to="loc_us-gaap_CommonStockMember_8ab80ab8-f2b6-4e25-8c20-5f38670a6386" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AdditionalPaidInCapitalMember_401729d2-c4a4-48fb-b36b-571691e42aa4" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AdditionalPaidInCapitalMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EquityComponentDomain_bc771a3a-8eb0-4205-acbc-c6f477c9e9ac" xlink:to="loc_us-gaap_AdditionalPaidInCapitalMember_401729d2-c4a4-48fb-b36b-571691e42aa4" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_RetainedEarningsMember_9b544394-5e89-461c-ad54-720e4e29be2b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_RetainedEarningsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EquityComponentDomain_bc771a3a-8eb0-4205-acbc-c6f477c9e9ac" xlink:to="loc_us-gaap_RetainedEarningsMember_9b544394-5e89-461c-ad54-720e4e29be2b" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccumulatedOtherComprehensiveIncomeMember_cf2b45c5-6890-4c76-8411-7b6208440957" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AccumulatedOtherComprehensiveIncomeMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EquityComponentDomain_bc771a3a-8eb0-4205-acbc-c6f477c9e9ac" xlink:to="loc_us-gaap_AccumulatedOtherComprehensiveIncomeMember_cf2b45c5-6890-4c76-8411-7b6208440957" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TreasuryStockMember_844ba9b0-76e6-4baf-8da9-dfdf071bc36d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TreasuryStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EquityComponentDomain_bc771a3a-8eb0-4205-acbc-c6f477c9e9ac" xlink:to="loc_us-gaap_TreasuryStockMember_844ba9b0-76e6-4baf-8da9-dfdf071bc36d" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TreasuryStockCommonMember_267323f9-3ca4-4ded-86ed-acb485e3d382" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TreasuryStockCommonMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EquityComponentDomain_bc771a3a-8eb0-4205-acbc-c6f477c9e9ac" xlink:to="loc_us-gaap_TreasuryStockCommonMember_267323f9-3ca4-4ded-86ed-acb485e3d382" xlink:type="arc" order="6"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/ConsolidatedStatementofCashFlows" xlink:type="simple" xlink:href="trup-20201231.xsd#ConsolidatedStatementofCashFlows"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ConsolidatedStatementofCashFlows" xlink:type="extended" id="i1bfe9ebfe37845d19a30ec105cfdab1a_ConsolidatedStatementofCashFlows"/>
<link:roleRef roleURI="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPolicies" xlink:type="simple" xlink:href="trup-20201231.xsd#NatureofOperationsandSummaryofSignificantAccountingPolicies"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPolicies" xlink:type="extended" id="i4460245541764a5591bfde4a7bbb97da_NatureofOperationsandSummaryofSignificantAccountingPolicies"/>
<link:roleRef roleURI="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPoliciesPolicies" xlink:type="simple" xlink:href="trup-20201231.xsd#NatureofOperationsandSummaryofSignificantAccountingPoliciesPolicies"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPoliciesPolicies" xlink:type="extended" id="i0963d183812c4d128ee9b160d0b7d1ab_NatureofOperationsandSummaryofSignificantAccountingPoliciesPolicies"/>
<link:roleRef roleURI="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPoliciesNarrativeDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#NatureofOperationsandSummaryofSignificantAccountingPoliciesNarrativeDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPoliciesNarrativeDetails" xlink:type="extended" id="i17d22a7fda8d495dbce717e6fdcbcd96_NatureofOperationsandSummaryofSignificantAccountingPoliciesNarrativeDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPoliciesDeferredAcquisitionCostsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#NatureofOperationsandSummaryofSignificantAccountingPoliciesDeferredAcquisitionCostsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPoliciesDeferredAcquisitionCostsDetails" xlink:type="extended" id="i201ab8eb63b148fa9515a66db0cd287d_NatureofOperationsandSummaryofSignificantAccountingPoliciesDeferredAcquisitionCostsDetails">
<link:loc xlink:type="locator" xlink:label="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersLineItems_13afd7bc-9949-4178-ae95-67770b28e9e4" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersAmortizationOfDeferredPolicyAcquisitionCosts_6a5f12d0-740c-4445-b172-246740658deb" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersAmortizationOfDeferredPolicyAcquisitionCosts"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersLineItems_13afd7bc-9949-4178-ae95-67770b28e9e4" xlink:to="loc_us-gaap_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersAmortizationOfDeferredPolicyAcquisitionCosts_6a5f12d0-740c-4445-b172-246740658deb" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersTable_3a7a9f07-22d4-4070-96d2-fd997a2f61b9" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersLineItems_13afd7bc-9949-4178-ae95-67770b28e9e4" xlink:to="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersTable_3a7a9f07-22d4-4070-96d2-fd997a2f61b9" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeStatementLocationAxis_ecbf855e-4bd6-4b2c-9750-847983bd5362" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeStatementLocationAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersTable_3a7a9f07-22d4-4070-96d2-fd997a2f61b9" xlink:to="loc_us-gaap_IncomeStatementLocationAxis_ecbf855e-4bd6-4b2c-9750-847983bd5362" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeStatementLocationDomain_ecbf855e-4bd6-4b2c-9750-847983bd5362_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeStatementLocationDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_IncomeStatementLocationAxis_ecbf855e-4bd6-4b2c-9750-847983bd5362" xlink:to="loc_us-gaap_IncomeStatementLocationDomain_ecbf855e-4bd6-4b2c-9750-847983bd5362_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeStatementLocationDomain_0a48388e-bf1b-4854-a2af-432e3b514da8" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeStatementLocationDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_IncomeStatementLocationAxis_ecbf855e-4bd6-4b2c-9750-847983bd5362" xlink:to="loc_us-gaap_IncomeStatementLocationDomain_0a48388e-bf1b-4854-a2af-432e3b514da8" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SellingAndMarketingExpenseMember_a6344a42-c0fc-47c4-ac39-4717afdec0cc" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SellingAndMarketingExpenseMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncomeStatementLocationDomain_0a48388e-bf1b-4854-a2af-432e3b514da8" xlink:to="loc_us-gaap_SellingAndMarketingExpenseMember_a6344a42-c0fc-47c4-ac39-4717afdec0cc" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CostOfSalesMember_c40860b1-858a-4247-95f8-a9235d1d4045" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CostOfSalesMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncomeStatementLocationDomain_0a48388e-bf1b-4854-a2af-432e3b514da8" xlink:to="loc_us-gaap_CostOfSalesMember_c40860b1-858a-4247-95f8-a9235d1d4045" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersByAffiliationWithRegistrantAxis_2c914f65-4cc5-4a37-b4b3-b0f8bf8a1361" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersByAffiliationWithRegistrantAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersTable_3a7a9f07-22d4-4070-96d2-fd997a2f61b9" xlink:to="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersByAffiliationWithRegistrantAxis_2c914f65-4cc5-4a37-b4b3-b0f8bf8a1361" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_PropertyCasualtyInsuranceUnderwritersAffiliationWithRegistrantDomain_2c914f65-4cc5-4a37-b4b3-b0f8bf8a1361_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_PropertyCasualtyInsuranceUnderwritersAffiliationWithRegistrantDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersByAffiliationWithRegistrantAxis_2c914f65-4cc5-4a37-b4b3-b0f8bf8a1361" xlink:to="loc_srt_PropertyCasualtyInsuranceUnderwritersAffiliationWithRegistrantDomain_2c914f65-4cc5-4a37-b4b3-b0f8bf8a1361_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_PropertyCasualtyInsuranceUnderwritersAffiliationWithRegistrantDomain_8b438058-5ad1-4b47-b07d-3b934a09c290" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_PropertyCasualtyInsuranceUnderwritersAffiliationWithRegistrantDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersByAffiliationWithRegistrantAxis_2c914f65-4cc5-4a37-b4b3-b0f8bf8a1361" xlink:to="loc_srt_PropertyCasualtyInsuranceUnderwritersAffiliationWithRegistrantDomain_8b438058-5ad1-4b47-b07d-3b934a09c290" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeAxis_1716eb8e-2151-47ab-8784-5538b4731f09" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_srt_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersTable_3a7a9f07-22d4-4070-96d2-fd997a2f61b9" xlink:to="loc_srt_RangeAxis_1716eb8e-2151-47ab-8784-5538b4731f09" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeMember_1716eb8e-2151-47ab-8784-5538b4731f09_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_RangeAxis_1716eb8e-2151-47ab-8784-5538b4731f09" xlink:to="loc_srt_RangeMember_1716eb8e-2151-47ab-8784-5538b4731f09_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeMember_3f11bcf5-6486-4f5d-b13e-3a87af59803c" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_RangeAxis_1716eb8e-2151-47ab-8784-5538b4731f09" xlink:to="loc_srt_RangeMember_3f11bcf5-6486-4f5d-b13e-3a87af59803c" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPoliciesPropertyPlantandEquipmentUsefulLifeDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#NatureofOperationsandSummaryofSignificantAccountingPoliciesPropertyPlantandEquipmentUsefulLifeDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/NatureofOperationsandSummaryofSignificantAccountingPoliciesPropertyPlantandEquipmentUsefulLifeDetails" xlink:type="extended" id="i60cb9c520ac84515a8e084a2e71a526a_NatureofOperationsandSummaryofSignificantAccountingPoliciesPropertyPlantandEquipmentUsefulLifeDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentLineItems_fa204f81-296f-47a2-86e0-36fd20a7a40b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentUsefulLife_dc7884ba-df88-46e5-81a7-8c472dff991c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentUsefulLife"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentLineItems_fa204f81-296f-47a2-86e0-36fd20a7a40b" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentUsefulLife_dc7884ba-df88-46e5-81a7-8c472dff991c" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ScheduleOfPropertyPlantAndEquipmentTable_62098c43-a139-4f74-9f28-8f0561cd8b4b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ScheduleOfPropertyPlantAndEquipmentTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentLineItems_fa204f81-296f-47a2-86e0-36fd20a7a40b" xlink:to="loc_us-gaap_ScheduleOfPropertyPlantAndEquipmentTable_62098c43-a139-4f74-9f28-8f0561cd8b4b" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentByTypeAxis_8f788fb9-9929-4a34-9bcc-afa8fbcc52c2" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentByTypeAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfPropertyPlantAndEquipmentTable_62098c43-a139-4f74-9f28-8f0561cd8b4b" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentByTypeAxis_8f788fb9-9929-4a34-9bcc-afa8fbcc52c2" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_8f788fb9-9929-4a34-9bcc-afa8fbcc52c2_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentTypeDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentByTypeAxis_8f788fb9-9929-4a34-9bcc-afa8fbcc52c2" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_8f788fb9-9929-4a34-9bcc-afa8fbcc52c2_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_6a7d8481-de3c-483f-a92f-242f0f9eddd4" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentTypeDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentByTypeAxis_8f788fb9-9929-4a34-9bcc-afa8fbcc52c2" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_6a7d8481-de3c-483f-a92f-242f0f9eddd4" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LandImprovementsMember_19883239-10d5-4c27-80ad-9aa98af0a0d4" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LandImprovementsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_6a7d8481-de3c-483f-a92f-242f0f9eddd4" xlink:to="loc_us-gaap_LandImprovementsMember_19883239-10d5-4c27-80ad-9aa98af0a0d4" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_BuildingMember_4d2164df-589b-4477-aab6-f088edcb636d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_BuildingMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_6a7d8481-de3c-483f-a92f-242f0f9eddd4" xlink:to="loc_us-gaap_BuildingMember_4d2164df-589b-4477-aab6-f088edcb636d" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SoftwareAndSoftwareDevelopmentCostsMember_576ca91b-235c-42ac-a473-885265cfc783" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SoftwareAndSoftwareDevelopmentCostsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_6a7d8481-de3c-483f-a92f-242f0f9eddd4" xlink:to="loc_us-gaap_SoftwareAndSoftwareDevelopmentCostsMember_576ca91b-235c-42ac-a473-885265cfc783" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OfficeEquipmentMember_fe82c0c1-e1ad-4b21-9be0-bd95c44b4c5f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OfficeEquipmentMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_6a7d8481-de3c-483f-a92f-242f0f9eddd4" xlink:to="loc_us-gaap_OfficeEquipmentMember_fe82c0c1-e1ad-4b21-9be0-bd95c44b4c5f" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeAxis_e344c788-5ef2-4869-ad7b-3d6567e445a3" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfPropertyPlantAndEquipmentTable_62098c43-a139-4f74-9f28-8f0561cd8b4b" xlink:to="loc_srt_RangeAxis_e344c788-5ef2-4869-ad7b-3d6567e445a3" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeMember_e344c788-5ef2-4869-ad7b-3d6567e445a3_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_RangeAxis_e344c788-5ef2-4869-ad7b-3d6567e445a3" xlink:to="loc_srt_RangeMember_e344c788-5ef2-4869-ad7b-3d6567e445a3_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeMember_ebd8c11c-d5a5-4872-9719-a2861e03d637" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_RangeAxis_e344c788-5ef2-4869-ad7b-3d6567e445a3" xlink:to="loc_srt_RangeMember_ebd8c11c-d5a5-4872-9719-a2861e03d637" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_MinimumMember_294516bf-e30d-4924-9d6b-cf24e7318307" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_MinimumMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_RangeMember_ebd8c11c-d5a5-4872-9719-a2861e03d637" xlink:to="loc_srt_MinimumMember_294516bf-e30d-4924-9d6b-cf24e7318307" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_MaximumMember_80113755-09b4-49d1-995d-152b8c6ecc58" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_MaximumMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_RangeMember_ebd8c11c-d5a5-4872-9719-a2861e03d637" xlink:to="loc_srt_MaximumMember_80113755-09b4-49d1-995d-152b8c6ecc58" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/NetLossperShare" xlink:type="simple" xlink:href="trup-20201231.xsd#NetLossperShare"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/NetLossperShare" xlink:type="extended" id="ib743ffd9c9164c37953329138cc5970e_NetLossperShare"/>
<link:roleRef roleURI="http://www.trupanion.com/role/NetLossperShareTables" xlink:type="simple" xlink:href="trup-20201231.xsd#NetLossperShareTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/NetLossperShareTables" xlink:type="extended" id="i961495538de14441911f39d0c64c4b59_NetLossperShareTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/NetLossperShareDetailsScheduleofAntidilutiveSecuritiesExcludedfromComputationofEarningsPerShare" xlink:type="simple" xlink:href="trup-20201231.xsd#NetLossperShareDetailsScheduleofAntidilutiveSecuritiesExcludedfromComputationofEarningsPerShare"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/NetLossperShareDetailsScheduleofAntidilutiveSecuritiesExcludedfromComputationofEarningsPerShare" xlink:type="extended" id="i17ae06727bc94778adff316e04e7616c_NetLossperShareDetailsScheduleofAntidilutiveSecuritiesExcludedfromComputationofEarningsPerShare">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems_69a50f76-77c9-4a9d-9e2e-95c26fbb2422" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsOutstandingNumber_edd0217b-36e2-4e55-92d4-cad65617e3fc" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsOutstandingNumber"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_AntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems_69a50f76-77c9-4a9d-9e2e-95c26fbb2422" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsOutstandingNumber_edd0217b-36e2-4e55-92d4-cad65617e3fc" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Restrictedstockoutstanding_39f7aef4-8e04-48ee-a197-648cd9949893" xlink:href="trup-20201231.xsd#trup_Restrictedstockoutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_AntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems_69a50f76-77c9-4a9d-9e2e-95c26fbb2422" xlink:to="loc_trup_Restrictedstockoutstanding_39f7aef4-8e04-48ee-a197-648cd9949893" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfWarrantOrRightOutstanding_e05407c7-33c4-46c7-8d2e-bc686f43c177" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfWarrantOrRightOutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_AntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems_69a50f76-77c9-4a9d-9e2e-95c26fbb2422" xlink:to="loc_us-gaap_ClassOfWarrantOrRightOutstanding_e05407c7-33c4-46c7-8d2e-bc686f43c177" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ScheduleOfAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareTable_369f88e7-7d9d-4dfe-b3cf-ac9accfad659" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ScheduleOfAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_AntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems_69a50f76-77c9-4a9d-9e2e-95c26fbb2422" xlink:to="loc_us-gaap_ScheduleOfAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareTable_369f88e7-7d9d-4dfe-b3cf-ac9accfad659" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementClassOfStockAxis_455de790-08eb-4792-8f2b-b28c5637fa81" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementClassOfStockAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareTable_369f88e7-7d9d-4dfe-b3cf-ac9accfad659" xlink:to="loc_us-gaap_StatementClassOfStockAxis_455de790-08eb-4792-8f2b-b28c5637fa81" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_455de790-08eb-4792-8f2b-b28c5637fa81_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementClassOfStockAxis_455de790-08eb-4792-8f2b-b28c5637fa81" xlink:to="loc_us-gaap_ClassOfStockDomain_455de790-08eb-4792-8f2b-b28c5637fa81_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_8a54bd08-d10d-41a8-a835-79660a819b0b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementClassOfStockAxis_455de790-08eb-4792-8f2b-b28c5637fa81" xlink:to="loc_us-gaap_ClassOfStockDomain_8a54bd08-d10d-41a8-a835-79660a819b0b" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StockOptionMember_c1d4ee43-7e9a-46f4-a35d-d200d321c087" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StockOptionMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_8a54bd08-d10d-41a8-a835-79660a819b0b" xlink:to="loc_us-gaap_StockOptionMember_c1d4ee43-7e9a-46f4-a35d-d200d321c087" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_RestrictedStockUnitsRSUMember_f7191f8a-a33b-449d-8ce3-970ff5b04ec7" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_RestrictedStockUnitsRSUMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_8a54bd08-d10d-41a8-a835-79660a819b0b" xlink:to="loc_us-gaap_RestrictedStockUnitsRSUMember_f7191f8a-a33b-449d-8ce3-970ff5b04ec7" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_WarrantMember_abee18c4-9979-4667-b2af-db83d1c33341" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_WarrantMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_8a54bd08-d10d-41a8-a835-79660a819b0b" xlink:to="loc_us-gaap_WarrantMember_abee18c4-9979-4667-b2af-db83d1c33341" xlink:type="arc" order="2"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/BusinessCombinations" xlink:type="simple" xlink:href="trup-20201231.xsd#BusinessCombinations"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/BusinessCombinations" xlink:type="extended" id="i8a12ecb9619149e0a94c209071925f0e_BusinessCombinations"/>
<link:roleRef roleURI="http://www.trupanion.com/role/BusinessCombinationsTables" xlink:type="simple" xlink:href="trup-20201231.xsd#BusinessCombinationsTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/BusinessCombinationsTables" xlink:type="extended" id="idd31567c006944309a16036018bcee85_BusinessCombinationsTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/BusinessCombinationsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#BusinessCombinationsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/BusinessCombinationsDetails" xlink:type="extended" id="ied5a1ca5cf25446bb2d78eeaf46f9a78_BusinessCombinationsDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/PropertyPlantandEquipmentNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#PropertyPlantandEquipmentNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/PropertyPlantandEquipmentNotes" xlink:type="extended" id="i621f480c013c4ff292a451d4ca273eab_PropertyPlantandEquipmentNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/PropertyPlantandEquipmentTables" xlink:type="simple" xlink:href="trup-20201231.xsd#PropertyPlantandEquipmentTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/PropertyPlantandEquipmentTables" xlink:type="extended" id="i4dee15f168814f468b9a20d49ac87a64_PropertyPlantandEquipmentTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/PropertyPlantandEquipmentDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#PropertyPlantandEquipmentDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/PropertyPlantandEquipmentDetails" xlink:type="extended" id="ic1a099bb93d14b66a3a55f3433d7aaa2_PropertyPlantandEquipmentDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentLineItems_ed923ec3-e5ee-4484-ba09-dfa2f8a32a36" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentGross_9729eb0d-aa7a-4f65-8533-70dae0a62d10" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentGross"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentLineItems_ed923ec3-e5ee-4484-ba09-dfa2f8a32a36" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentGross_9729eb0d-aa7a-4f65-8533-70dae0a62d10" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ConstructionInProgressGross_eccd2e61-368e-475f-bf64-543b4d57571d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ConstructionInProgressGross"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentLineItems_ed923ec3-e5ee-4484-ba09-dfa2f8a32a36" xlink:to="loc_us-gaap_ConstructionInProgressGross_eccd2e61-368e-475f-bf64-543b4d57571d" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccumulatedDepreciationDepletionAndAmortizationPropertyPlantAndEquipment_28f87095-3269-4089-ab9b-8e59c6f844cb" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AccumulatedDepreciationDepletionAndAmortizationPropertyPlantAndEquipment"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentLineItems_ed923ec3-e5ee-4484-ba09-dfa2f8a32a36" xlink:to="loc_us-gaap_AccumulatedDepreciationDepletionAndAmortizationPropertyPlantAndEquipment_28f87095-3269-4089-ab9b-8e59c6f844cb" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentNet_366f931a-1dfc-4e1e-b94d-0b32aa30b535" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentLineItems_ed923ec3-e5ee-4484-ba09-dfa2f8a32a36" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentNet_366f931a-1dfc-4e1e-b94d-0b32aa30b535" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_Depreciation_b9df9054-2986-4098-bb40-dad4ca0841e6" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_Depreciation"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentLineItems_ed923ec3-e5ee-4484-ba09-dfa2f8a32a36" xlink:to="loc_us-gaap_Depreciation_b9df9054-2986-4098-bb40-dad4ca0841e6" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ScheduleOfPropertyPlantAndEquipmentTable_831a558e-a02e-452c-b8b3-6b4877e1075d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ScheduleOfPropertyPlantAndEquipmentTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentLineItems_ed923ec3-e5ee-4484-ba09-dfa2f8a32a36" xlink:to="loc_us-gaap_ScheduleOfPropertyPlantAndEquipmentTable_831a558e-a02e-452c-b8b3-6b4877e1075d" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeAxis_7a7cd779-5b9e-4e44-8abc-752e3b333155" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfPropertyPlantAndEquipmentTable_831a558e-a02e-452c-b8b3-6b4877e1075d" xlink:to="loc_srt_RangeAxis_7a7cd779-5b9e-4e44-8abc-752e3b333155" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeMember_7a7cd779-5b9e-4e44-8abc-752e3b333155_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_RangeAxis_7a7cd779-5b9e-4e44-8abc-752e3b333155" xlink:to="loc_srt_RangeMember_7a7cd779-5b9e-4e44-8abc-752e3b333155_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeMember_52ae3d0e-2a10-4656-8034-6653b8297710" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_RangeAxis_7a7cd779-5b9e-4e44-8abc-752e3b333155" xlink:to="loc_srt_RangeMember_52ae3d0e-2a10-4656-8034-6653b8297710" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentByTypeAxis_9f3f6fa5-1f07-4c0a-b996-3c5d79539cf8" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentByTypeAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfPropertyPlantAndEquipmentTable_831a558e-a02e-452c-b8b3-6b4877e1075d" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentByTypeAxis_9f3f6fa5-1f07-4c0a-b996-3c5d79539cf8" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_9f3f6fa5-1f07-4c0a-b996-3c5d79539cf8_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentTypeDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentByTypeAxis_9f3f6fa5-1f07-4c0a-b996-3c5d79539cf8" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_9f3f6fa5-1f07-4c0a-b996-3c5d79539cf8_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_374f559d-a46c-4de0-b989-c438a74f3b20" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentTypeDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentByTypeAxis_9f3f6fa5-1f07-4c0a-b996-3c5d79539cf8" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_374f559d-a46c-4de0-b989-c438a74f3b20" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LandAndLandImprovementsMember_55a79e5c-5fd0-4630-9cea-4d2902558d7b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LandAndLandImprovementsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_374f559d-a46c-4de0-b989-c438a74f3b20" xlink:to="loc_us-gaap_LandAndLandImprovementsMember_55a79e5c-5fd0-4630-9cea-4d2902558d7b" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_BuildingAndBuildingImprovementsMember_2587b4e3-2702-4cb8-9077-b67907052467" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_BuildingAndBuildingImprovementsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_374f559d-a46c-4de0-b989-c438a74f3b20" xlink:to="loc_us-gaap_BuildingAndBuildingImprovementsMember_2587b4e3-2702-4cb8-9077-b67907052467" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SoftwareAndSoftwareDevelopmentCostsMember_93d25183-fb30-4a73-bec7-da3763c6ff76" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SoftwareAndSoftwareDevelopmentCostsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_374f559d-a46c-4de0-b989-c438a74f3b20" xlink:to="loc_us-gaap_SoftwareAndSoftwareDevelopmentCostsMember_93d25183-fb30-4a73-bec7-da3763c6ff76" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ComputerEquipmentMember_b4d3c957-d2cf-44fa-bf4c-cec5d93bcdef" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ComputerEquipmentMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentTypeDomain_374f559d-a46c-4de0-b989-c438a74f3b20" xlink:to="loc_us-gaap_ComputerEquipmentMember_b4d3c957-d2cf-44fa-bf4c-cec5d93bcdef" xlink:type="arc" order="3"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/PropertyPlantandEquipmentRealEstateAcquisitionDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#PropertyPlantandEquipmentRealEstateAcquisitionDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/PropertyPlantandEquipmentRealEstateAcquisitionDetails" xlink:type="extended" id="i2352a3ae4d6441b988dfcd5358c4554e_PropertyPlantandEquipmentRealEstateAcquisitionDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IntangibleAssetsNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#IntangibleAssetsNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IntangibleAssetsNotes" xlink:type="extended" id="i179cbb878d6e45f2add638a03b6ca2f5_IntangibleAssetsNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IntangibleAssetsTables" xlink:type="simple" xlink:href="trup-20201231.xsd#IntangibleAssetsTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IntangibleAssetsTables" xlink:type="extended" id="i791027d54e9d4b35925ea128d1b89287_IntangibleAssetsTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IntangibleAssetsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#IntangibleAssetsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IntangibleAssetsDetails" xlink:type="extended" id="i93194315496640e1b9544190f968b756_IntangibleAssetsDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseNextTwelveMonths_4e39b03c-0d19-42d5-8de0-463020a0360d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseNextTwelveMonths"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseNextTwelveMonths_4e39b03c-0d19-42d5-8de0-463020a0360d" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IndefiniteLivedLicenseAgreements_8965b7d4-f41b-43b3-a9e9-9e53ed65f8ee" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IndefiniteLivedLicenseAgreements"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_IndefiniteLivedLicenseAgreements_8965b7d4-f41b-43b3-a9e9-9e53ed65f8ee" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedPatentsGross_e27a9509-5186-4ac8-ad56-004e717f276a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedPatentsGross"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_FiniteLivedPatentsGross_e27a9509-5186-4ac8-ad56-004e717f276a" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IntangibleAssetsNetExcludingGoodwill_839c8bcf-9344-4b05-96f8-69a00a9c0f88" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IntangibleAssetsNetExcludingGoodwill"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_IntangibleAssetsNetExcludingGoodwill_839c8bcf-9344-4b05-96f8-69a00a9c0f88" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsAccumulatedAmortization_1c742bd1-f28c-4aff-9047-8d8a9c6a149e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsAccumulatedAmortization"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsAccumulatedAmortization_1c742bd1-f28c-4aff-9047-8d8a9c6a149e" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearTwo_d6e0c43d-758c-48ee-b293-e96a13a37d1e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearTwo"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearTwo_d6e0c43d-758c-48ee-b293-e96a13a37d1e" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearThree_0c0de06b-e480-4beb-8c50-6a4cc5e9c519" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearThree"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearThree_0c0de06b-e480-4beb-8c50-6a4cc5e9c519" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearFour_711a04fe-713f-4a75-a338-155414417a29" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearFour"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearFour_711a04fe-713f-4a75-a338-155414417a29" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearFive_6c55bdec-fc40-43bb-b502-de1a9cb666fc" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearFive"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseYearFive_6c55bdec-fc40-43bb-b502-de1a9cb666fc" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseAfterYearFive_3e5cceb6-e2f5-469d-a47b-b5cf946a5746" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseAfterYearFive"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsAmortizationExpenseAfterYearFive_3e5cceb6-e2f5-469d-a47b-b5cf946a5746" xlink:type="arc" order="9"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_FiniteLivedIntangibleAssetsTotalExpectedAmortizationExpense_14b4e2c4-5847-4ad8-90a1-6c353470e905" xlink:href="trup-20201231.xsd#trup_FiniteLivedIntangibleAssetsTotalExpectedAmortizationExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_trup_FiniteLivedIntangibleAssetsTotalExpectedAmortizationExpense_14b4e2c4-5847-4ad8-90a1-6c353470e905" xlink:type="arc" order="10"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Leaserelatedintangibleassetsweightedaverageusefullife_0ab9dcb9-7f35-476a-9529-5bc1d8aaec36" xlink:href="trup-20201231.xsd#trup_Leaserelatedintangibleassetsweightedaverageusefullife"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_trup_Leaserelatedintangibleassetsweightedaverageusefullife_0ab9dcb9-7f35-476a-9529-5bc1d8aaec36" xlink:type="arc" order="11"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AcquiredFiniteLivedIntangibleAssetsWeightedAverageUsefulLife_d12bcb31-feb1-4483-b9eb-45b6cde2fb8d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AcquiredFiniteLivedIntangibleAssetsWeightedAverageUsefulLife"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_AcquiredFiniteLivedIntangibleAssetsWeightedAverageUsefulLife_d12bcb31-feb1-4483-b9eb-45b6cde2fb8d" xlink:type="arc" order="12"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_GoodwillAcquiredDuringPeriod_de1fae33-4c1c-42e6-84fb-9ba771f64c21" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_GoodwillAcquiredDuringPeriod"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_GoodwillAcquiredDuringPeriod_de1fae33-4c1c-42e6-84fb-9ba771f64c21" xlink:type="arc" order="13"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_Goodwill_3f0e3465-1a14-4c43-83f0-2cadbaa44b67" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_Goodwill"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_Goodwill_3f0e3465-1a14-4c43-83f0-2cadbaa44b67" xlink:type="arc" order="14"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_GoodwillForeignCurrencyTranslationGainLoss_822a1671-25c5-46dd-858b-180b9a6b9571" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_GoodwillForeignCurrencyTranslationGainLoss"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_GoodwillForeignCurrencyTranslationGainLoss_822a1671-25c5-46dd-858b-180b9a6b9571" xlink:type="arc" order="15"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ScheduleOfFiniteLivedIntangibleAssetsTable_e0275321-cb0e-4394-8ec8-f93a326b34c2" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ScheduleOfFiniteLivedIntangibleAssetsTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsLineItems_b357fd8a-9110-4f47-8bdb-b8c4654b2477" xlink:to="loc_us-gaap_ScheduleOfFiniteLivedIntangibleAssetsTable_e0275321-cb0e-4394-8ec8-f93a326b34c2" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsByMajorClassAxis_854b269e-4dde-4011-b0bf-1825224a7cae" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsByMajorClassAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfFiniteLivedIntangibleAssetsTable_e0275321-cb0e-4394-8ec8-f93a326b34c2" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsByMajorClassAxis_854b269e-4dde-4011-b0bf-1825224a7cae" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain_854b269e-4dde-4011-b0bf-1825224a7cae_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsByMajorClassAxis_854b269e-4dde-4011-b0bf-1825224a7cae" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain_854b269e-4dde-4011-b0bf-1825224a7cae_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain_150fb0e4-cb73-48c3-a1d8-bdd39792b1a9" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsByMajorClassAxis_854b269e-4dde-4011-b0bf-1825224a7cae" xlink:to="loc_us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain_150fb0e4-cb73-48c3-a1d8-bdd39792b1a9" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PatentsMember_f058bfa2-995a-452f-85e2-1fafda3249b7" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PatentsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain_150fb0e4-cb73-48c3-a1d8-bdd39792b1a9" xlink:to="loc_us-gaap_PatentsMember_f058bfa2-995a-452f-85e2-1fafda3249b7" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LeasesAcquiredInPlaceMember_66763202-1e12-4169-9a67-def237ff2372" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LeasesAcquiredInPlaceMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain_150fb0e4-cb73-48c3-a1d8-bdd39792b1a9" xlink:to="loc_us-gaap_LeasesAcquiredInPlaceMember_66763202-1e12-4169-9a67-def237ff2372" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TradeNamesMember_e774fcb0-83ef-416b-ba3e-c0b1002d6392" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TradeNamesMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain_150fb0e4-cb73-48c3-a1d8-bdd39792b1a9" xlink:to="loc_us-gaap_TradeNamesMember_e774fcb0-83ef-416b-ba3e-c0b1002d6392" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TechnologyBasedIntangibleAssetsMember_bba7fd34-ae23-4e9a-a754-038cf18ca562" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TechnologyBasedIntangibleAssetsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain_150fb0e4-cb73-48c3-a1d8-bdd39792b1a9" xlink:to="loc_us-gaap_TechnologyBasedIntangibleAssetsMember_bba7fd34-ae23-4e9a-a754-038cf18ca562" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CustomerRelationshipsMember_04ec2782-2c75-4b20-a33f-b2f6edb69158" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CustomerRelationshipsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FiniteLivedIntangibleAssetsMajorClassNameDomain_150fb0e4-cb73-48c3-a1d8-bdd39792b1a9" xlink:to="loc_us-gaap_CustomerRelationshipsMember_04ec2782-2c75-4b20-a33f-b2f6edb69158" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidationItemsAxis_3a049ea9-0b3c-4a48-bc5e-0a29143480bc" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidationItemsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfFiniteLivedIntangibleAssetsTable_e0275321-cb0e-4394-8ec8-f93a326b34c2" xlink:to="loc_srt_ConsolidationItemsAxis_3a049ea9-0b3c-4a48-bc5e-0a29143480bc" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidationItemsDomain_3a049ea9-0b3c-4a48-bc5e-0a29143480bc_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidationItemsDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_ConsolidationItemsAxis_3a049ea9-0b3c-4a48-bc5e-0a29143480bc" xlink:to="loc_srt_ConsolidationItemsDomain_3a049ea9-0b3c-4a48-bc5e-0a29143480bc_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidationItemsDomain_3d8b5e77-e25f-4acf-a2f0-980a59fd2860" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidationItemsDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_ConsolidationItemsAxis_3a049ea9-0b3c-4a48-bc5e-0a29143480bc" xlink:to="loc_srt_ConsolidationItemsDomain_3d8b5e77-e25f-4acf-a2f0-980a59fd2860" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementBusinessSegmentsAxis_a849b014-498a-43c8-816d-c07d70f3bf3c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementBusinessSegmentsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfFiniteLivedIntangibleAssetsTable_e0275321-cb0e-4394-8ec8-f93a326b34c2" xlink:to="loc_us-gaap_StatementBusinessSegmentsAxis_a849b014-498a-43c8-816d-c07d70f3bf3c" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_a849b014-498a-43c8-816d-c07d70f3bf3c_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_a849b014-498a-43c8-816d-c07d70f3bf3c" xlink:to="loc_us-gaap_SegmentDomain_a849b014-498a-43c8-816d-c07d70f3bf3c_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_efa4e8b7-c827-427b-a342-05a2f951fb43" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_a849b014-498a-43c8-816d-c07d70f3bf3c" xlink:to="loc_us-gaap_SegmentDomain_efa4e8b7-c827-427b-a342-05a2f951fb43" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeAxis_b18246fb-a35b-4c8b-8820-0c02db60ac2e" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfFiniteLivedIntangibleAssetsTable_e0275321-cb0e-4394-8ec8-f93a326b34c2" xlink:to="loc_srt_RangeAxis_b18246fb-a35b-4c8b-8820-0c02db60ac2e" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeMember_b18246fb-a35b-4c8b-8820-0c02db60ac2e_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_RangeAxis_b18246fb-a35b-4c8b-8820-0c02db60ac2e" xlink:to="loc_srt_RangeMember_b18246fb-a35b-4c8b-8820-0c02db60ac2e_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_RangeMember_b585b41e-dfee-4c60-bbb1-5afc1da8ea57" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_RangeMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_RangeAxis_b18246fb-a35b-4c8b-8820-0c02db60ac2e" xlink:to="loc_srt_RangeMember_b585b41e-dfee-4c60-bbb1-5afc1da8ea57" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/InvestmentSecuritiesNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#InvestmentSecuritiesNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/InvestmentSecuritiesNotes" xlink:type="extended" id="i8bc1aadb6e4248af86841cd77e2796a5_InvestmentSecuritiesNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/InvestmentSecuritiesAvailableforSaleTables" xlink:type="simple" xlink:href="trup-20201231.xsd#InvestmentSecuritiesAvailableforSaleTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/InvestmentSecuritiesAvailableforSaleTables" xlink:type="extended" id="icbfcf6c851bd471685dfd2b11626f7b6_InvestmentSecuritiesAvailableforSaleTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/InvestmentSecuritiesDetailsInvestmentSchedule" xlink:type="simple" xlink:href="trup-20201231.xsd#InvestmentSecuritiesDetailsInvestmentSchedule"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/InvestmentSecuritiesDetailsInvestmentSchedule" xlink:type="extended" id="ia66da25f149a4144a630a51289324dae_InvestmentSecuritiesDetailsInvestmentSchedule">
<link:loc xlink:type="locator" xlink:label="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:href="trup-20201231.xsd#trup_InvestmentLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AvailableForSaleSecuritiesAmortizedCost_0c1f458d-ff1d-4f07-8818-2bfabdfab8b7" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AvailableForSaleSecuritiesAmortizedCost"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:to="loc_us-gaap_AvailableForSaleSecuritiesAmortizedCost_0c1f458d-ff1d-4f07-8818-2bfabdfab8b7" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AvailableForSaleSecuritiesAccumulatedGrossUnrealizedGainBeforeTax_496e83a3-d038-45e2-a071-d20941bb0fbd" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AvailableForSaleSecuritiesAccumulatedGrossUnrealizedGainBeforeTax"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:to="loc_us-gaap_AvailableForSaleSecuritiesAccumulatedGrossUnrealizedGainBeforeTax_496e83a3-d038-45e2-a071-d20941bb0fbd" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AvailableForSaleSecuritiesAccumulatedGrossUnrealizedLossBeforeTax_096c0011-3df8-42c5-8edf-934b2bf73970" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AvailableForSaleSecuritiesAccumulatedGrossUnrealizedLossBeforeTax"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:to="loc_us-gaap_AvailableForSaleSecuritiesAccumulatedGrossUnrealizedLossBeforeTax_096c0011-3df8-42c5-8edf-934b2bf73970" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AvailableForSaleSecuritiesDebtMaturitiesWithoutSingleMaturityDateFairValue_c7dc474d-7803-47f3-912a-6472b7431d24" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AvailableForSaleSecuritiesDebtMaturitiesWithoutSingleMaturityDateFairValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:to="loc_us-gaap_AvailableForSaleSecuritiesDebtMaturitiesWithoutSingleMaturityDateFairValue_c7dc474d-7803-47f3-912a-6472b7431d24" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_HeldToMaturitySecuritiesAmortizedCostBeforeOtherThanTemporaryImpairment_2884e75a-9e68-4c64-b6e7-54f9866dbc4f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_HeldToMaturitySecuritiesAmortizedCostBeforeOtherThanTemporaryImpairment"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:to="loc_us-gaap_HeldToMaturitySecuritiesAmortizedCostBeforeOtherThanTemporaryImpairment_2884e75a-9e68-4c64-b6e7-54f9866dbc4f" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_HeldToMaturitySecuritiesAccumulatedUnrecognizedHoldingGain_d9b0f85d-8b6b-443d-9f99-a7c8f47a0fb6" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_HeldToMaturitySecuritiesAccumulatedUnrecognizedHoldingGain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:to="loc_us-gaap_HeldToMaturitySecuritiesAccumulatedUnrecognizedHoldingGain_d9b0f85d-8b6b-443d-9f99-a7c8f47a0fb6" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_HeldToMaturitySecuritiesAccumulatedUnrecognizedHoldingLoss_15a2c066-a918-4380-89fa-588d34d32939" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_HeldToMaturitySecuritiesAccumulatedUnrecognizedHoldingLoss"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:to="loc_us-gaap_HeldToMaturitySecuritiesAccumulatedUnrecognizedHoldingLoss_15a2c066-a918-4380-89fa-588d34d32939" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_HeldToMaturitySecuritiesFairValue_965cc5d9-44c3-45b7-90cf-d4fb38d08beb" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_HeldToMaturitySecuritiesFairValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:to="loc_us-gaap_HeldToMaturitySecuritiesFairValue_965cc5d9-44c3-45b7-90cf-d4fb38d08beb" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_InvestmentTable_290c20b1-482e-44aa-9477-72f2e6e1466a" xlink:href="trup-20201231.xsd#trup_InvestmentTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_trup_InvestmentLineItems_332b9f5e-7c83-4013-928a-c32ed3256904" xlink:to="loc_trup_InvestmentTable_290c20b1-482e-44aa-9477-72f2e6e1466a" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_InvestmentTypeAxis_f033fb78-d5fa-4fa8-9929-96d66d1883c4" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_InvestmentTypeAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_trup_InvestmentTable_290c20b1-482e-44aa-9477-72f2e6e1466a" xlink:to="loc_us-gaap_InvestmentTypeAxis_f033fb78-d5fa-4fa8-9929-96d66d1883c4" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_InvestmentTypeCategorizationMember_f033fb78-d5fa-4fa8-9929-96d66d1883c4_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_InvestmentTypeCategorizationMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_InvestmentTypeAxis_f033fb78-d5fa-4fa8-9929-96d66d1883c4" xlink:to="loc_us-gaap_InvestmentTypeCategorizationMember_f033fb78-d5fa-4fa8-9929-96d66d1883c4_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_InvestmentTypeCategorizationMember_4747f4d3-dfc3-4e4d-a2c5-41f8104a9264" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_InvestmentTypeCategorizationMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_InvestmentTypeAxis_f033fb78-d5fa-4fa8-9929-96d66d1883c4" xlink:to="loc_us-gaap_InvestmentTypeCategorizationMember_4747f4d3-dfc3-4e4d-a2c5-41f8104a9264" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_DepositsMember_21b7e7f7-8e35-47d7-a0d5-153391e04655" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_DepositsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_InvestmentTypeCategorizationMember_4747f4d3-dfc3-4e4d-a2c5-41f8104a9264" xlink:to="loc_us-gaap_DepositsMember_21b7e7f7-8e35-47d7-a0d5-153391e04655" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_MunicipalBondsMember_30e21be6-3ee2-4c8c-bb6d-1a75a040297a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_MunicipalBondsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_InvestmentTypeCategorizationMember_4747f4d3-dfc3-4e4d-a2c5-41f8104a9264" xlink:to="loc_us-gaap_MunicipalBondsMember_30e21be6-3ee2-4c8c-bb6d-1a75a040297a" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_USTreasurySecuritiesMember_08f48491-df85-4ad0-a88d-c8e0b031396d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_USTreasurySecuritiesMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_InvestmentTypeCategorizationMember_4747f4d3-dfc3-4e4d-a2c5-41f8104a9264" xlink:to="loc_us-gaap_USTreasurySecuritiesMember_08f48491-df85-4ad0-a88d-c8e0b031396d" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CertificatesOfDepositMember_a1a18b96-688c-4294-bdd9-7462dee295b2" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CertificatesOfDepositMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_InvestmentTypeCategorizationMember_4747f4d3-dfc3-4e4d-a2c5-41f8104a9264" xlink:to="loc_us-gaap_CertificatesOfDepositMember_a1a18b96-688c-4294-bdd9-7462dee295b2" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_USGovernmentDebtSecuritiesMember_ae2d5778-09e7-4311-8ebb-f0467cc242db" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_USGovernmentDebtSecuritiesMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_InvestmentTypeCategorizationMember_4747f4d3-dfc3-4e4d-a2c5-41f8104a9264" xlink:to="loc_us-gaap_USGovernmentDebtSecuritiesMember_ae2d5778-09e7-4311-8ebb-f0467cc242db" xlink:type="arc" order="4"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/InvestmentSecuritiesDetailsAvailableforSale" xlink:type="simple" xlink:href="trup-20201231.xsd#InvestmentSecuritiesDetailsAvailableforSale"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/InvestmentSecuritiesDetailsAvailableforSale" xlink:type="extended" id="i8fa4c3513b8b45ad9cefaaa99fe3a1a3_InvestmentSecuritiesDetailsAvailableforSale"/>
<link:roleRef roleURI="http://www.trupanion.com/role/OtherInvestmentsNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#OtherInvestmentsNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/OtherInvestmentsNotes" xlink:type="extended" id="i1cfb5b6e94324b0a9ce8bd760889041e_OtherInvestmentsNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/OtherInvestmentsdetailsPolicies" xlink:type="simple" xlink:href="trup-20201231.xsd#OtherInvestmentsdetailsPolicies"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/OtherInvestmentsdetailsPolicies" xlink:type="extended" id="i8ae3b293e90b4e01a7fb85a3e324fccd_OtherInvestmentsdetailsPolicies"/>
<link:roleRef roleURI="http://www.trupanion.com/role/OtherInvestmentsTables" xlink:type="simple" xlink:href="trup-20201231.xsd#OtherInvestmentsTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/OtherInvestmentsTables" xlink:type="extended" id="iafd2955ff05c49f485e09c411a76c4ca_OtherInvestmentsTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/OtherInvestmentsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#OtherInvestmentsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/OtherInvestmentsDetails" xlink:type="extended" id="i9a3327f2e6a04f0a9527f55fe5e99de6_OtherInvestmentsDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/FairValue" xlink:type="simple" xlink:href="trup-20201231.xsd#FairValue"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/FairValue" xlink:type="extended" id="ia2fd06a9cf1d4998bf5d985c3bb1cb65_FairValue"/>
<link:roleRef roleURI="http://www.trupanion.com/role/FairValueTables" xlink:type="simple" xlink:href="trup-20201231.xsd#FairValueTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/FairValueTables" xlink:type="extended" id="i7bb306bfeb134385b5b2fd2b58ae1288_FairValueTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/FairValueDetailsUnobservable" xlink:type="simple" xlink:href="trup-20201231.xsd#FairValueDetailsUnobservable"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/FairValueDetailsUnobservable" xlink:type="extended" id="ib85b1dda358a44e5bbbc7462abba773a_FairValueDetailsUnobservable">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems_fe4c0ac2-e3b2-4414-8360-b6df23e0a953" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_RestrictedCashAndCashEquivalentsNoncurrent_3221f2ea-fe83-4bc2-80d4-84cbd9ec0b2d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_RestrictedCashAndCashEquivalentsNoncurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems_fe4c0ac2-e3b2-4414-8360-b6df23e0a953" xlink:to="loc_us-gaap_RestrictedCashAndCashEquivalentsNoncurrent_3221f2ea-fe83-4bc2-80d4-84cbd9ec0b2d" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_MarketableSecuritiesNoncurrent_08f07b30-0714-4d34-9b1e-b2ed92dad589" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_MarketableSecuritiesNoncurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems_fe4c0ac2-e3b2-4414-8360-b6df23e0a953" xlink:to="loc_us-gaap_MarketableSecuritiesNoncurrent_08f07b30-0714-4d34-9b1e-b2ed92dad589" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CashAndCashEquivalentsFairValueDisclosure_0e78e479-5982-4d3d-86ac-6c9d4a387548" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CashAndCashEquivalentsFairValueDisclosure"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems_fe4c0ac2-e3b2-4414-8360-b6df23e0a953" xlink:to="loc_us-gaap_CashAndCashEquivalentsFairValueDisclosure_0e78e479-5982-4d3d-86ac-6c9d4a387548" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AssetsFairValueDisclosure_93eea44a-c8ca-4ac9-bbf6-044574815c73" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AssetsFairValueDisclosure"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems_fe4c0ac2-e3b2-4414-8360-b6df23e0a953" xlink:to="loc_us-gaap_AssetsFairValueDisclosure_93eea44a-c8ca-4ac9-bbf6-044574815c73" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_NoncontrollingInterestInVariableInterestEntity_24b4bc34-be42-4976-b29a-036eb19c218d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_NoncontrollingInterestInVariableInterestEntity"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems_fe4c0ac2-e3b2-4414-8360-b6df23e0a953" xlink:to="loc_us-gaap_NoncontrollingInterestInVariableInterestEntity_24b4bc34-be42-4976-b29a-036eb19c218d" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_NotesReceivableFairValueDisclosure_eab3d423-7ab6-47e4-a4dc-3813bcc5d880" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_NotesReceivableFairValueDisclosure"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems_fe4c0ac2-e3b2-4414-8360-b6df23e0a953" xlink:to="loc_us-gaap_NotesReceivableFairValueDisclosure_eab3d423-7ab6-47e4-a4dc-3813bcc5d880" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherComprehensiveIncomeUnrealizedHoldingGainLossOnSecuritiesArisingDuringPeriodNetOfTax_5fa50a33-be74-4167-a5fa-7185c368aaaf" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherComprehensiveIncomeUnrealizedHoldingGainLossOnSecuritiesArisingDuringPeriodNetOfTax"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems_fe4c0ac2-e3b2-4414-8360-b6df23e0a953" xlink:to="loc_us-gaap_OtherComprehensiveIncomeUnrealizedHoldingGainLossOnSecuritiesArisingDuringPeriodNetOfTax_5fa50a33-be74-4167-a5fa-7185c368aaaf" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisTable_592551f8-e5a4-45a0-99ca-1be828ccf38a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisLineItems_fe4c0ac2-e3b2-4414-8360-b6df23e0a953" xlink:to="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisTable_592551f8-e5a4-45a0-99ca-1be828ccf38a" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_InvestmentTypeAxis_ded62936-05e0-41a5-9a69-b2257eebbe16" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_InvestmentTypeAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisTable_592551f8-e5a4-45a0-99ca-1be828ccf38a" xlink:to="loc_us-gaap_InvestmentTypeAxis_ded62936-05e0-41a5-9a69-b2257eebbe16" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_InvestmentTypeCategorizationMember_ded62936-05e0-41a5-9a69-b2257eebbe16_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_InvestmentTypeCategorizationMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_InvestmentTypeAxis_ded62936-05e0-41a5-9a69-b2257eebbe16" xlink:to="loc_us-gaap_InvestmentTypeCategorizationMember_ded62936-05e0-41a5-9a69-b2257eebbe16_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_InvestmentTypeCategorizationMember_721ab7ad-b7c6-4d0f-b5a1-75c26f5171c1" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_InvestmentTypeCategorizationMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_InvestmentTypeAxis_ded62936-05e0-41a5-9a69-b2257eebbe16" xlink:to="loc_us-gaap_InvestmentTypeCategorizationMember_721ab7ad-b7c6-4d0f-b5a1-75c26f5171c1" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CashAndCashEquivalentsMember_ed11a579-af41-4673-a7be-795b6ef55bf5" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CashAndCashEquivalentsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_InvestmentTypeCategorizationMember_721ab7ad-b7c6-4d0f-b5a1-75c26f5171c1" xlink:to="loc_us-gaap_CashAndCashEquivalentsMember_ed11a579-af41-4673-a7be-795b6ef55bf5" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_DepositsMember_b2d67d97-6f4f-4e65-9627-5d0edea22548" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_DepositsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_InvestmentTypeCategorizationMember_721ab7ad-b7c6-4d0f-b5a1-75c26f5171c1" xlink:to="loc_us-gaap_DepositsMember_b2d67d97-6f4f-4e65-9627-5d0edea22548" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_MunicipalBondsMember_ecf998d9-e85b-4eda-befc-2e15249dd429" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_MunicipalBondsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_InvestmentTypeCategorizationMember_721ab7ad-b7c6-4d0f-b5a1-75c26f5171c1" xlink:to="loc_us-gaap_MunicipalBondsMember_ecf998d9-e85b-4eda-befc-2e15249dd429" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_MoneyMarketFundsMember_1219c89b-aaaa-4ce7-8f72-0f763352b8f0" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_MoneyMarketFundsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_InvestmentTypeCategorizationMember_721ab7ad-b7c6-4d0f-b5a1-75c26f5171c1" xlink:to="loc_us-gaap_MoneyMarketFundsMember_1219c89b-aaaa-4ce7-8f72-0f763352b8f0" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FairValueByFairValueHierarchyLevelAxis_f77b8556-5ed4-440f-aa88-fd0092851608" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FairValueByFairValueHierarchyLevelAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_FairValueAssetsAndLiabilitiesMeasuredOnRecurringAndNonrecurringBasisTable_592551f8-e5a4-45a0-99ca-1be828ccf38a" xlink:to="loc_us-gaap_FairValueByFairValueHierarchyLevelAxis_f77b8556-5ed4-440f-aa88-fd0092851608" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FairValueMeasurementsFairValueHierarchyDomain_f77b8556-5ed4-440f-aa88-fd0092851608_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FairValueMeasurementsFairValueHierarchyDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_FairValueByFairValueHierarchyLevelAxis_f77b8556-5ed4-440f-aa88-fd0092851608" xlink:to="loc_us-gaap_FairValueMeasurementsFairValueHierarchyDomain_f77b8556-5ed4-440f-aa88-fd0092851608_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FairValueMeasurementsFairValueHierarchyDomain_3e91a5d4-7967-452d-9062-c1c72c545b44" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FairValueMeasurementsFairValueHierarchyDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_FairValueByFairValueHierarchyLevelAxis_f77b8556-5ed4-440f-aa88-fd0092851608" xlink:to="loc_us-gaap_FairValueMeasurementsFairValueHierarchyDomain_3e91a5d4-7967-452d-9062-c1c72c545b44" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FairValueInputsLevel1Member_cf1fd344-cc99-40e7-85b0-ab08a844a667" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FairValueInputsLevel1Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueMeasurementsFairValueHierarchyDomain_3e91a5d4-7967-452d-9062-c1c72c545b44" xlink:to="loc_us-gaap_FairValueInputsLevel1Member_cf1fd344-cc99-40e7-85b0-ab08a844a667" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FairValueInputsLevel2Member_2b3053e0-ae58-41a3-b852-195bc1c1d619" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FairValueInputsLevel2Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueMeasurementsFairValueHierarchyDomain_3e91a5d4-7967-452d-9062-c1c72c545b44" xlink:to="loc_us-gaap_FairValueInputsLevel2Member_2b3053e0-ae58-41a3-b852-195bc1c1d619" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_FairValueInputsLevel3Member_81937356-0630-4c46-8399-b4f22d485b83" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_FairValueInputsLevel3Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_FairValueMeasurementsFairValueHierarchyDomain_3e91a5d4-7967-452d-9062-c1c72c545b44" xlink:to="loc_us-gaap_FairValueInputsLevel3Member_81937356-0630-4c46-8399-b4f22d485b83" xlink:type="arc" order="2"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/CommitmentandContingencies" xlink:type="simple" xlink:href="trup-20201231.xsd#CommitmentandContingencies"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/CommitmentandContingencies" xlink:type="extended" id="i89a9600032c244fba886b4b50cd307ac_CommitmentandContingencies"/>
<link:roleRef roleURI="http://www.trupanion.com/role/CommitmentandContingenciesTables" xlink:type="simple" xlink:href="trup-20201231.xsd#CommitmentandContingenciesTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/CommitmentandContingenciesTables" xlink:type="extended" id="ie55ca3f92698499e9930fe09cb5f506f_CommitmentandContingenciesTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/CommitmentandContingenciesDetailsNarrative" xlink:type="simple" xlink:href="trup-20201231.xsd#CommitmentandContingenciesDetailsNarrative"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/CommitmentandContingenciesDetailsNarrative" xlink:type="extended" id="i20eb75209a4f470387d8468c2a7c1e2e_CommitmentandContingenciesDetailsNarrative"/>
<link:roleRef roleURI="http://www.trupanion.com/role/CommitmentandContingenciesCommitmentsandContingenciesDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#CommitmentandContingenciesCommitmentsandContingenciesDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/CommitmentandContingenciesCommitmentsandContingenciesDetails" xlink:type="extended" id="i2376bbeae9e84749a4a81c40bf23bef7_CommitmentandContingenciesCommitmentsandContingenciesDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentsLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentDueInNextTwelveMonths_a22f668a-8aa3-460e-8775-089d52ad157c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentDueInNextTwelveMonths"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OtherCommitmentDueInNextTwelveMonths_a22f668a-8aa3-460e-8775-089d52ad157c" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentDueInSecondYear_f3f39178-7a58-4617-a4bf-ceb0536f13e8" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentDueInSecondYear"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OtherCommitmentDueInSecondYear_f3f39178-7a58-4617-a4bf-ceb0536f13e8" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentDueInThirdYear_ef5e00cc-fecb-40c2-b3be-1ab4225b77cb" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentDueInThirdYear"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OtherCommitmentDueInThirdYear_ef5e00cc-fecb-40c2-b3be-1ab4225b77cb" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentDueInFourthYear_f077e46d-92c4-4508-aa29-5ab37cefa877" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentDueInFourthYear"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OtherCommitmentDueInFourthYear_f077e46d-92c4-4508-aa29-5ab37cefa877" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentDueInFifthYear_fba4d27b-61a1-4317-ae08-f257a0e22c26" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentDueInFifthYear"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OtherCommitmentDueInFifthYear_fba4d27b-61a1-4317-ae08-f257a0e22c26" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentDueAfterFifthYear_3ccb0315-75d2-4bb5-9666-1ae8ac98b5e6" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentDueAfterFifthYear"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OtherCommitmentDueAfterFifthYear_3ccb0315-75d2-4bb5-9666-1ae8ac98b5e6" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitment_14d338db-b6d0-41eb-aa7f-d7c0b33e417e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitment"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OtherCommitment_14d338db-b6d0-41eb-aa7f-d7c0b33e417e" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Totalfuturecommitmentsdueinnexttwelvemonths_93690a5f-b58a-4c7a-ae17-323ac4989d08" xlink:href="trup-20201231.xsd#trup_Totalfuturecommitmentsdueinnexttwelvemonths"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_trup_Totalfuturecommitmentsdueinnexttwelvemonths_93690a5f-b58a-4c7a-ae17-323ac4989d08" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Totalfuturecommitmentsdueintwoyears_a2bfc5db-cfb1-4bb3-a035-26b0e2a57fa5" xlink:href="trup-20201231.xsd#trup_Totalfuturecommitmentsdueintwoyears"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_trup_Totalfuturecommitmentsdueintwoyears_a2bfc5db-cfb1-4bb3-a035-26b0e2a57fa5" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Totalfuturecommitmentsdueinthreeyears_9ea22d26-3968-43d7-8bb7-fc676a5eabc9" xlink:href="trup-20201231.xsd#trup_Totalfuturecommitmentsdueinthreeyears"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_trup_Totalfuturecommitmentsdueinthreeyears_9ea22d26-3968-43d7-8bb7-fc676a5eabc9" xlink:type="arc" order="9"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Totalfuturecommitmentsdueinfouryears_d0ef0a77-2842-49f4-aeb7-4a8bf440de0e" xlink:href="trup-20201231.xsd#trup_Totalfuturecommitmentsdueinfouryears"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_trup_Totalfuturecommitmentsdueinfouryears_d0ef0a77-2842-49f4-aeb7-4a8bf440de0e" xlink:type="arc" order="10"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Totalfuturecommitmentsdueinfiveyears_d4a15674-ac64-48be-968d-12c30c473ee1" xlink:href="trup-20201231.xsd#trup_Totalfuturecommitmentsdueinfiveyears"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_trup_Totalfuturecommitmentsdueinfiveyears_d4a15674-ac64-48be-968d-12c30c473ee1" xlink:type="arc" order="11"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Totalfuturecommitmentsduethereafter_7bafa442-59c4-402f-8dce-50820992a4c3" xlink:href="trup-20201231.xsd#trup_Totalfuturecommitmentsduethereafter"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_trup_Totalfuturecommitmentsduethereafter_7bafa442-59c4-402f-8dce-50820992a4c3" xlink:type="arc" order="12"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Totalfuturecommitmentspaymentsdue_daa7f075-d056-4859-b122-44b112c03386" xlink:href="trup-20201231.xsd#trup_Totalfuturecommitmentspaymentsdue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_trup_Totalfuturecommitmentspaymentsdue_daa7f075-d056-4859-b122-44b112c03386" xlink:type="arc" order="13"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueCurrent_31f46c4c-c519-4175-8475-4a8c6226847f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OperatingLeasesFutureMinimumPaymentsDueCurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueCurrent_31f46c4c-c519-4175-8475-4a8c6226847f" xlink:type="arc" order="14"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueInTwoYears_26d4802b-653b-47a7-a3c6-9abd86df39e7" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OperatingLeasesFutureMinimumPaymentsDueInTwoYears"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueInTwoYears_26d4802b-653b-47a7-a3c6-9abd86df39e7" xlink:type="arc" order="15"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueInThreeYears_e1e027e9-2865-40f6-b4fc-e5a6d068d5f3" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OperatingLeasesFutureMinimumPaymentsDueInThreeYears"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueInThreeYears_e1e027e9-2865-40f6-b4fc-e5a6d068d5f3" xlink:type="arc" order="16"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueInFourYears_49eacc4b-255f-452e-a0d6-e83989feef2a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OperatingLeasesFutureMinimumPaymentsDueInFourYears"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueInFourYears_49eacc4b-255f-452e-a0d6-e83989feef2a" xlink:type="arc" order="17"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueInFiveYears_9f5f3169-8fac-4ef6-b643-dd652e7a0a71" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OperatingLeasesFutureMinimumPaymentsDueInFiveYears"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueInFiveYears_9f5f3169-8fac-4ef6-b643-dd652e7a0a71" xlink:type="arc" order="18"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueThereafter_f50c871b-9be4-4e03-9565-5e923b6197d6" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OperatingLeasesFutureMinimumPaymentsDueThereafter"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDueThereafter_f50c871b-9be4-4e03-9565-5e923b6197d6" xlink:type="arc" order="19"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDue_6e084cc4-2a92-4e55-9e9a-20709ce8b11e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OperatingLeasesFutureMinimumPaymentsDue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OperatingLeasesFutureMinimumPaymentsDue_6e084cc4-2a92-4e55-9e9a-20709ce8b11e" xlink:type="arc" order="20"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentsTable_e00e1217-0a6a-4efe-9c5f-258c78ca09c8" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentsTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_OtherCommitmentsLineItems_27348e26-921d-41c8-8d07-88499592666e" xlink:to="loc_us-gaap_OtherCommitmentsTable_e00e1217-0a6a-4efe-9c5f-258c78ca09c8" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentsAxis_0f9f14a7-321a-486b-a1b0-339ef907e267" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_OtherCommitmentsTable_e00e1217-0a6a-4efe-9c5f-258c78ca09c8" xlink:to="loc_us-gaap_OtherCommitmentsAxis_0f9f14a7-321a-486b-a1b0-339ef907e267" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentsDomain_0f9f14a7-321a-486b-a1b0-339ef907e267_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentsDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_OtherCommitmentsAxis_0f9f14a7-321a-486b-a1b0-339ef907e267" xlink:to="loc_us-gaap_OtherCommitmentsDomain_0f9f14a7-321a-486b-a1b0-339ef907e267_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCommitmentsDomain_0d9f4303-c17e-46bd-857c-56de0a4871c9" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCommitmentsDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_OtherCommitmentsAxis_0f9f14a7-321a-486b-a1b0-339ef907e267" xlink:to="loc_us-gaap_OtherCommitmentsDomain_0d9f4303-c17e-46bd-857c-56de0a4871c9" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/ClaimsReserveNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#ClaimsReserveNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ClaimsReserveNotes" xlink:type="extended" id="i761ccce04f0d48168ec79ba9a728ffa1_ClaimsReserveNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/ClaimsReserveTables" xlink:type="simple" xlink:href="trup-20201231.xsd#ClaimsReserveTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ClaimsReserveTables" xlink:type="extended" id="i763a9fdcaaed460ba11050782c47aa7e_ClaimsReserveTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/ClaimsReserveDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#ClaimsReserveDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ClaimsReserveDetails" xlink:type="extended" id="i553f01a9af9b404085f23b81031fca3d_ClaimsReserveDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_ef7ac474-90cc-418d-b6d4-6e7d4a0a69bd" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_ChangeinLiabilityforUnpaidClaimsandClaimsAdjustmentExpenseNet_85ed4297-04f6-4d6e-bcf7-107d87f9c066" xlink:href="trup-20201231.xsd#trup_ChangeinLiabilityforUnpaidClaimsandClaimsAdjustmentExpenseNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_ef7ac474-90cc-418d-b6d4-6e7d4a0a69bd" xlink:to="loc_trup_ChangeinLiabilityforUnpaidClaimsandClaimsAdjustmentExpenseNet_85ed4297-04f6-4d6e-bcf7-107d87f9c066" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsCumulativePaidClaimsAndAllocatedClaimAdjustmentExpenseNet_2f2bf8cf-cb05-4645-8246-721f160641cf" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsCumulativePaidClaimsAndAllocatedClaimAdjustmentExpenseNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_ef7ac474-90cc-418d-b6d4-6e7d4a0a69bd" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsCumulativePaidClaimsAndAllocatedClaimAdjustmentExpenseNet_2f2bf8cf-cb05-4645-8246-721f160641cf" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsIncurredClaimsAndAllocatedClaimAdjustmentExpenseNet_f808cc00-0266-4394-9fd5-1510f70060b3" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsIncurredClaimsAndAllocatedClaimAdjustmentExpenseNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_ef7ac474-90cc-418d-b6d4-6e7d4a0a69bd" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsIncurredClaimsAndAllocatedClaimAdjustmentExpenseNet_f808cc00-0266-4394-9fd5-1510f70060b3" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsIncurredButNotReportedIbnrClaimsLiabilityNet_43257ac4-d2bd-4aa3-8978-99ebfa795698" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsIncurredButNotReportedIbnrClaimsLiabilityNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_ef7ac474-90cc-418d-b6d4-6e7d4a0a69bd" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsIncurredButNotReportedIbnrClaimsLiabilityNet_43257ac4-d2bd-4aa3-8978-99ebfa795698" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsNumberOfReportedClaims_6f592747-45f9-41ab-89d2-2a5d2a7196f6" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsNumberOfReportedClaims"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_ef7ac474-90cc-418d-b6d4-6e7d4a0a69bd" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsNumberOfReportedClaims_6f592747-45f9-41ab-89d2-2a5d2a7196f6" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_ef7ac474-90cc-418d-b6d4-6e7d4a0a69bd" xlink:to="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseNet_f3883488-709a-4e55-9915-0b2944b76cea" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:to="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseNet_f3883488-709a-4e55-9915-0b2944b76cea" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersCurrentYearClaimsAndClaimsAdjustmentExpense_85819e49-d646-4da9-8e70-f9bfeca1a06a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersCurrentYearClaimsAndClaimsAdjustmentExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:to="loc_us-gaap_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersCurrentYearClaimsAndClaimsAdjustmentExpense_85819e49-d646-4da9-8e70-f9bfeca1a06a" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersPriorYearClaimsAndClaimsAdjustmentExpense_86256468-5d03-4578-b54e-3482eadf3166" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersPriorYearClaimsAndClaimsAdjustmentExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:to="loc_us-gaap_SupplementalInformationForPropertyCasualtyInsuranceUnderwritersPriorYearClaimsAndClaimsAdjustmentExpense_86256468-5d03-4578-b54e-3482eadf3166" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseClaimsPaidCurrentYear1_ca5d4d54-f6c8-4d16-a703-dffadb6e5616" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseClaimsPaidCurrentYear1"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:to="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseClaimsPaidCurrentYear1_ca5d4d54-f6c8-4d16-a703-dffadb6e5616" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseClaimsPaidPriorYears1_0714c168-6b3d-4ca6-ba27-3a82fadadd42" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseClaimsPaidPriorYears1"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:to="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseClaimsPaidPriorYears1_0714c168-6b3d-4ca6-ba27-3a82fadadd42" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PaymentsForLossesAndLossAdjustmentExpense_3f4b03eb-4a88-462f-971a-ee9067713b4e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PaymentsForLossesAndLossAdjustmentExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:to="loc_us-gaap_PaymentsForLossesAndLossAdjustmentExpense_3f4b03eb-4a88-462f-971a-ee9067713b4e" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Claimsexpensenoncash_cf04380e-d8f8-4a18-b4f3-1eea9e114332" xlink:href="trup-20201231.xsd#trup_Claimsexpensenoncash"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:to="loc_trup_Claimsexpensenoncash_cf04380e-d8f8-4a18-b4f3-1eea9e114332" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseIncurredClaims1_ca493234-2a17-4cd5-8b72-9427265a0a6d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseIncurredClaims1"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseAbstract_45236620-4f7b-4481-a034-6f3e8e5dd945" xlink:to="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseIncurredClaims1_ca493234-2a17-4cd5-8b72-9427265a0a6d" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseNet_9c61c649-415c-4395-ae4c-8255f788f2cf" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseNet"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable_f4e5650f-614e-459a-ac55-82d5f8c3f6e3" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_ef7ac474-90cc-418d-b6d4-6e7d4a0a69bd" xlink:to="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable_f4e5650f-614e-459a-ac55-82d5f8c3f6e3" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementBusinessSegmentsAxis_b9aa2030-4699-4098-b144-1e97d8f2c664" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementBusinessSegmentsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable_f4e5650f-614e-459a-ac55-82d5f8c3f6e3" xlink:to="loc_us-gaap_StatementBusinessSegmentsAxis_b9aa2030-4699-4098-b144-1e97d8f2c664" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_b9aa2030-4699-4098-b144-1e97d8f2c664_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_b9aa2030-4699-4098-b144-1e97d8f2c664" xlink:to="loc_us-gaap_SegmentDomain_b9aa2030-4699-4098-b144-1e97d8f2c664_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_659535cb-b3e5-454e-a230-babd73b29a84" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_b9aa2030-4699-4098-b144-1e97d8f2c664" xlink:to="loc_us-gaap_SegmentDomain_659535cb-b3e5-454e-a230-babd73b29a84" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_SubscriptionbusinessMember_43f2d128-ac4c-4439-b703-ce71b1c1a5a6" xlink:href="trup-20201231.xsd#trup_SubscriptionbusinessMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_659535cb-b3e5-454e-a230-babd73b29a84" xlink:to="loc_trup_SubscriptionbusinessMember_43f2d128-ac4c-4439-b703-ce71b1c1a5a6" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AllOtherSegmentsMember_0533841b-15ba-4078-a4f7-8be6fc6d0b2d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AllOtherSegmentsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_659535cb-b3e5-454e-a230-babd73b29a84" xlink:to="loc_us-gaap_AllOtherSegmentsMember_0533841b-15ba-4078-a4f7-8be6fc6d0b2d" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/ClaimsReserveCumulativeclaimspaidandclaimsadjustmentexpensesDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#ClaimsReserveCumulativeclaimspaidandclaimsadjustmentexpensesDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ClaimsReserveCumulativeclaimspaidandclaimsadjustmentexpensesDetails" xlink:type="extended" id="ic846778f690d4cd2a172376375eb91e3_ClaimsReserveCumulativeclaimspaidandclaimsadjustmentexpensesDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClaimsDevelopmentLineItems_5f9185d3-5722-44e1-87e6-dec88465c293" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClaimsDevelopmentLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsNumberOfReportedClaims_7e479b83-bab7-4c22-b0cf-ac28e1bc9c0e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsNumberOfReportedClaims"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClaimsDevelopmentLineItems_5f9185d3-5722-44e1-87e6-dec88465c293" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsNumberOfReportedClaims_7e479b83-bab7-4c22-b0cf-ac28e1bc9c0e" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsIncurredClaimsAndAllocatedClaimAdjustmentExpenseNet_4cedd1a8-6d4b-4392-899c-40bfdfaccb77" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsIncurredClaimsAndAllocatedClaimAdjustmentExpenseNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClaimsDevelopmentLineItems_5f9185d3-5722-44e1-87e6-dec88465c293" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsIncurredClaimsAndAllocatedClaimAdjustmentExpenseNet_4cedd1a8-6d4b-4392-899c-40bfdfaccb77" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsIncurredButNotReportedIbnrClaimsLiabilityNet_1d436d74-30bf-4d22-9cc5-904fbcbfe2d3" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsIncurredButNotReportedIbnrClaimsLiabilityNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClaimsDevelopmentLineItems_5f9185d3-5722-44e1-87e6-dec88465c293" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsIncurredButNotReportedIbnrClaimsLiabilityNet_1d436d74-30bf-4d22-9cc5-904fbcbfe2d3" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsClaimsDevelopmentTable_52405457-072c-477c-859e-63bc998ecfed" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsClaimsDevelopmentTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_ClaimsDevelopmentLineItems_5f9185d3-5722-44e1-87e6-dec88465c293" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsClaimsDevelopmentTable_52405457-072c-477c-859e-63bc998ecfed" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ProductOrServiceAxis_374814f5-ec41-4c10-b33f-ed05855bce02" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ProductOrServiceAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsClaimsDevelopmentTable_52405457-072c-477c-859e-63bc998ecfed" xlink:to="loc_srt_ProductOrServiceAxis_374814f5-ec41-4c10-b33f-ed05855bce02" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ProductsAndServicesDomain_374814f5-ec41-4c10-b33f-ed05855bce02_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ProductsAndServicesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_ProductOrServiceAxis_374814f5-ec41-4c10-b33f-ed05855bce02" xlink:to="loc_srt_ProductsAndServicesDomain_374814f5-ec41-4c10-b33f-ed05855bce02_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ProductsAndServicesDomain_adcc4af7-4ae9-45ab-8a3c-19e55d186b2b" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ProductsAndServicesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_ProductOrServiceAxis_374814f5-ec41-4c10-b33f-ed05855bce02" xlink:to="loc_srt_ProductsAndServicesDomain_adcc4af7-4ae9-45ab-8a3c-19e55d186b2b" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_StatementGeographicalAxis_7ce2a8fd-28d8-4243-9b7d-946e81d736b7" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_StatementGeographicalAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsClaimsDevelopmentTable_52405457-072c-477c-859e-63bc998ecfed" xlink:to="loc_srt_StatementGeographicalAxis_7ce2a8fd-28d8-4243-9b7d-946e81d736b7" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_SegmentGeographicalDomain_7ce2a8fd-28d8-4243-9b7d-946e81d736b7_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_SegmentGeographicalDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_StatementGeographicalAxis_7ce2a8fd-28d8-4243-9b7d-946e81d736b7" xlink:to="loc_srt_SegmentGeographicalDomain_7ce2a8fd-28d8-4243-9b7d-946e81d736b7_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_SegmentGeographicalDomain_515fdd2a-34f7-4357-84e4-e92bb4dd55a2" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_SegmentGeographicalDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_StatementGeographicalAxis_7ce2a8fd-28d8-4243-9b7d-946e81d736b7" xlink:to="loc_srt_SegmentGeographicalDomain_515fdd2a-34f7-4357-84e4-e92bb4dd55a2" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementBusinessSegmentsAxis_d661c7c0-5501-4a9c-b988-009c3f7f72f2" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementBusinessSegmentsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsClaimsDevelopmentTable_52405457-072c-477c-859e-63bc998ecfed" xlink:to="loc_us-gaap_StatementBusinessSegmentsAxis_d661c7c0-5501-4a9c-b988-009c3f7f72f2" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_d661c7c0-5501-4a9c-b988-009c3f7f72f2_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_d661c7c0-5501-4a9c-b988-009c3f7f72f2" xlink:to="loc_us-gaap_SegmentDomain_d661c7c0-5501-4a9c-b988-009c3f7f72f2_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_08804db8-3763-461a-8ccd-b33667965644" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_d661c7c0-5501-4a9c-b988-009c3f7f72f2" xlink:to="loc_us-gaap_SegmentDomain_08804db8-3763-461a-8ccd-b33667965644" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AllOtherSegmentsMember_2df98f12-bc27-431d-9784-8baddc0d3f2d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AllOtherSegmentsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_08804db8-3763-461a-8ccd-b33667965644" xlink:to="loc_us-gaap_AllOtherSegmentsMember_2df98f12-bc27-431d-9784-8baddc0d3f2d" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_SubscriptionbusinessMember_6d351f0c-b8de-48e6-8b0b-31b9b9591748" xlink:href="trup-20201231.xsd#trup_SubscriptionbusinessMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_08804db8-3763-461a-8ccd-b33667965644" xlink:to="loc_trup_SubscriptionbusinessMember_6d351f0c-b8de-48e6-8b0b-31b9b9591748" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_MajorCustomersAxis_03d0c033-484a-4e8f-bb9b-279c2345e3c6" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_MajorCustomersAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsClaimsDevelopmentTable_52405457-072c-477c-859e-63bc998ecfed" xlink:to="loc_srt_MajorCustomersAxis_03d0c033-484a-4e8f-bb9b-279c2345e3c6" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_NameOfMajorCustomerDomain_03d0c033-484a-4e8f-bb9b-279c2345e3c6_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_NameOfMajorCustomerDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_MajorCustomersAxis_03d0c033-484a-4e8f-bb9b-279c2345e3c6" xlink:to="loc_srt_NameOfMajorCustomerDomain_03d0c033-484a-4e8f-bb9b-279c2345e3c6_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_NameOfMajorCustomerDomain_466acf95-fcf9-464a-aaed-4a35854d4c99" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_NameOfMajorCustomerDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_MajorCustomersAxis_03d0c033-484a-4e8f-bb9b-279c2345e3c6" xlink:to="loc_srt_NameOfMajorCustomerDomain_466acf95-fcf9-464a-aaed-4a35854d4c99" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearAxis_ce55dad1-fa95-41e8-a0f2-dc01b9748a28" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsAccidentYearAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsClaimsDevelopmentTable_52405457-072c-477c-859e-63bc998ecfed" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearAxis_ce55dad1-fa95-41e8-a0f2-dc01b9748a28" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_ce55dad1-fa95-41e8-a0f2-dc01b9748a28_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsAccidentYearDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearAxis_ce55dad1-fa95-41e8-a0f2-dc01b9748a28" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_ce55dad1-fa95-41e8-a0f2-dc01b9748a28_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_cef99832-b841-468d-8345-9078910ef1c9" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsAccidentYearDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearAxis_ce55dad1-fa95-41e8-a0f2-dc01b9748a28" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_cef99832-b841-468d-8345-9078910ef1c9" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortDurationInsuranceContractsAccidentYear2017Member_0a638eb2-839c-44f0-8a57-6c9f31d4d92a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortDurationInsuranceContractsAccidentYear2017Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_cef99832-b841-468d-8345-9078910ef1c9" xlink:to="loc_us-gaap_ShortDurationInsuranceContractsAccidentYear2017Member_0a638eb2-839c-44f0-8a57-6c9f31d4d92a" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsAccidentYear2016Member_25504b2e-bdf6-4ef6-af1e-002859fde60d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsAccidentYear2016Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_cef99832-b841-468d-8345-9078910ef1c9" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsAccidentYear2016Member_25504b2e-bdf6-4ef6-af1e-002859fde60d" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_ShortdurationInsuranceContractsAccidentYear2019Member_09476168-cf74-4ed5-8c75-6440f09643dc" xlink:href="trup-20201231.xsd#trup_ShortdurationInsuranceContractsAccidentYear2019Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_cef99832-b841-468d-8345-9078910ef1c9" xlink:to="loc_trup_ShortdurationInsuranceContractsAccidentYear2019Member_09476168-cf74-4ed5-8c75-6440f09643dc" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortDurationInsuranceContractsAccidentYear2018Member_a77f3fca-ec74-41b9-9cfd-b70626809330" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortDurationInsuranceContractsAccidentYear2018Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_cef99832-b841-468d-8345-9078910ef1c9" xlink:to="loc_us-gaap_ShortDurationInsuranceContractsAccidentYear2018Member_a77f3fca-ec74-41b9-9cfd-b70626809330" xlink:type="arc" order="3"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/ClaimsReserveIncurredclaimsandclaimadjustmentexpenseDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#ClaimsReserveIncurredclaimsandclaimadjustmentexpenseDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/ClaimsReserveIncurredclaimsandclaimadjustmentexpenseDetails" xlink:type="extended" id="iea6a086b88724529b8674af9ca4e1013_ClaimsReserveIncurredclaimsandclaimadjustmentexpenseDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_d3f877df-2c95-4179-8f6a-249c484c523f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsCumulativePaidClaimsAndAllocatedClaimAdjustmentExpenseNet_cb77b7a8-8930-4309-aa1f-e22c0e44bd12" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsCumulativePaidClaimsAndAllocatedClaimAdjustmentExpenseNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_d3f877df-2c95-4179-8f6a-249c484c523f" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsCumulativePaidClaimsAndAllocatedClaimAdjustmentExpenseNet_cb77b7a8-8930-4309-aa1f-e22c0e44bd12" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseNet_d0bd9039-9858-4f23-8525-59477a746186" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_d3f877df-2c95-4179-8f6a-249c484c523f" xlink:to="loc_us-gaap_LiabilityForUnpaidClaimsAndClaimsAdjustmentExpenseNet_d0bd9039-9858-4f23-8525-59477a746186" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable_3bc4de5d-75f7-4b9c-ae25-b2f67deb875e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseLineItems_d3f877df-2c95-4179-8f6a-249c484c523f" xlink:to="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable_3bc4de5d-75f7-4b9c-ae25-b2f67deb875e" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementBusinessSegmentsAxis_5b1e973c-25a1-464d-b895-f792d0c48daa" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementBusinessSegmentsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable_3bc4de5d-75f7-4b9c-ae25-b2f67deb875e" xlink:to="loc_us-gaap_StatementBusinessSegmentsAxis_5b1e973c-25a1-464d-b895-f792d0c48daa" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_5b1e973c-25a1-464d-b895-f792d0c48daa_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_5b1e973c-25a1-464d-b895-f792d0c48daa" xlink:to="loc_us-gaap_SegmentDomain_5b1e973c-25a1-464d-b895-f792d0c48daa_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_2e2e13f8-221b-4ad9-93b7-aac0d80138e8" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_5b1e973c-25a1-464d-b895-f792d0c48daa" xlink:to="loc_us-gaap_SegmentDomain_2e2e13f8-221b-4ad9-93b7-aac0d80138e8" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AllOtherSegmentsMember_28efab15-061d-47e5-a934-ec2afc6ac554" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AllOtherSegmentsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_2e2e13f8-221b-4ad9-93b7-aac0d80138e8" xlink:to="loc_us-gaap_AllOtherSegmentsMember_28efab15-061d-47e5-a934-ec2afc6ac554" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_SubscriptionbusinessMember_1dcd9427-e658-4461-805b-2bf2e52c27fb" xlink:href="trup-20201231.xsd#trup_SubscriptionbusinessMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_2e2e13f8-221b-4ad9-93b7-aac0d80138e8" xlink:to="loc_trup_SubscriptionbusinessMember_1dcd9427-e658-4461-805b-2bf2e52c27fb" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ProductOrServiceAxis_745c1338-13da-48c3-9b40-3f28aefdc7fe" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ProductOrServiceAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable_3bc4de5d-75f7-4b9c-ae25-b2f67deb875e" xlink:to="loc_srt_ProductOrServiceAxis_745c1338-13da-48c3-9b40-3f28aefdc7fe" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ProductsAndServicesDomain_745c1338-13da-48c3-9b40-3f28aefdc7fe_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ProductsAndServicesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_ProductOrServiceAxis_745c1338-13da-48c3-9b40-3f28aefdc7fe" xlink:to="loc_srt_ProductsAndServicesDomain_745c1338-13da-48c3-9b40-3f28aefdc7fe_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ProductsAndServicesDomain_4717defa-5de5-412e-8fa7-cd51aa95c78f" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ProductsAndServicesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_ProductOrServiceAxis_745c1338-13da-48c3-9b40-3f28aefdc7fe" xlink:to="loc_srt_ProductsAndServicesDomain_4717defa-5de5-412e-8fa7-cd51aa95c78f" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearAxis_6a124a11-8312-4c21-9822-2bc5ce0b47ad" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsAccidentYearAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_LiabilityForClaimsAndClaimsAdjustmentExpenseTable_3bc4de5d-75f7-4b9c-ae25-b2f67deb875e" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearAxis_6a124a11-8312-4c21-9822-2bc5ce0b47ad" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_6a124a11-8312-4c21-9822-2bc5ce0b47ad_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsAccidentYearDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearAxis_6a124a11-8312-4c21-9822-2bc5ce0b47ad" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_6a124a11-8312-4c21-9822-2bc5ce0b47ad_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_c80c343f-da6a-4dfd-9ebb-f169167137f2" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsAccidentYearDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearAxis_6a124a11-8312-4c21-9822-2bc5ce0b47ad" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_c80c343f-da6a-4dfd-9ebb-f169167137f2" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortDurationInsuranceContractsAccidentYear2018Member_33d75bde-4d46-40af-b703-721438ec0066" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortDurationInsuranceContractsAccidentYear2018Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_c80c343f-da6a-4dfd-9ebb-f169167137f2" xlink:to="loc_us-gaap_ShortDurationInsuranceContractsAccidentYear2018Member_33d75bde-4d46-40af-b703-721438ec0066" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortdurationInsuranceContractsAccidentYear2016Member_0697eaa9-ab58-49c6-8737-3eeeb156f514" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortdurationInsuranceContractsAccidentYear2016Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_c80c343f-da6a-4dfd-9ebb-f169167137f2" xlink:to="loc_us-gaap_ShortdurationInsuranceContractsAccidentYear2016Member_0697eaa9-ab58-49c6-8737-3eeeb156f514" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortDurationInsuranceContractsAccidentYear2017Member_e4855249-12b5-4d30-ba56-9709b96c89dc" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShortDurationInsuranceContractsAccidentYear2017Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_c80c343f-da6a-4dfd-9ebb-f169167137f2" xlink:to="loc_us-gaap_ShortDurationInsuranceContractsAccidentYear2017Member_e4855249-12b5-4d30-ba56-9709b96c89dc" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_ShortdurationInsuranceContractsAccidentYear2019Member_225ff95a-76ec-405c-bb23-6f7d65442c36" xlink:href="trup-20201231.xsd#trup_ShortdurationInsuranceContractsAccidentYear2019Member"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShortdurationInsuranceContractsAccidentYearDomain_c80c343f-da6a-4dfd-9ebb-f169167137f2" xlink:to="loc_trup_ShortdurationInsuranceContractsAccidentYear2019Member_225ff95a-76ec-405c-bb23-6f7d65442c36" xlink:type="arc" order="3"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/Debt" xlink:type="simple" xlink:href="trup-20201231.xsd#Debt"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/Debt" xlink:type="extended" id="ic45ed9135cb74c20b401b731ad11cc25_Debt"/>
<link:roleRef roleURI="http://www.trupanion.com/role/DebtDetailsNarrative" xlink:type="simple" xlink:href="trup-20201231.xsd#DebtDetailsNarrative"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/DebtDetailsNarrative" xlink:type="extended" id="i293138bfda894016890a5fd277882d79_DebtDetailsNarrative"/>
<link:roleRef roleURI="http://www.trupanion.com/role/StockbasedCompensation" xlink:type="simple" xlink:href="trup-20201231.xsd#StockbasedCompensation"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockbasedCompensation" xlink:type="extended" id="ice2079e6e27d40a5b0662d2ad6a165e0_StockbasedCompensation"/>
<link:roleRef roleURI="http://www.trupanion.com/role/StockbasedCompensationTables" xlink:type="simple" xlink:href="trup-20201231.xsd#StockbasedCompensationTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockbasedCompensationTables" xlink:type="extended" id="id074d90481a3445894fcfc66af78cecd_StockbasedCompensationTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/StockbasedCompensationDetailsNarrative" xlink:type="simple" xlink:href="trup-20201231.xsd#StockbasedCompensationDetailsNarrative"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockbasedCompensationDetailsNarrative" xlink:type="extended" id="i3aaacfeaf3294643a5a8d9205b2b47a7_StockbasedCompensationDetailsNarrative">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_10c03073-b5ce-4a89-885d-4642fad84f63" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardOptionsVestedInPeriodFairValue1_1f334853-31ec-44f0-9731-253db3d2b279" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardOptionsVestedInPeriodFairValue1"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_10c03073-b5ce-4a89-885d-4642fad84f63" xlink:to="loc_us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardOptionsVestedInPeriodFairValue1_1f334853-31ec-44f0-9731-253db3d2b279" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardOptionsNonvestedNumberOfShares_e18cd14c-f970-4235-88ab-9be943f24fa4" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardOptionsNonvestedNumberOfShares"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_10c03073-b5ce-4a89-885d-4642fad84f63" xlink:to="loc_us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardOptionsNonvestedNumberOfShares_e18cd14c-f970-4235-88ab-9be943f24fa4" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Restrictedstockoutstanding_126368a2-623e-4ec4-906a-3d83fa200f03" xlink:href="trup-20201231.xsd#trup_Restrictedstockoutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_10c03073-b5ce-4a89-885d-4642fad84f63" xlink:to="loc_trup_Restrictedstockoutstanding_126368a2-623e-4ec4-906a-3d83fa200f03" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_EmployeeServiceShareBasedCompensationNonvestedAwardsTotalCompensationCostNotYetRecognized_a1cda173-475b-49f6-a4fd-a03ac2d8f983" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_EmployeeServiceShareBasedCompensationNonvestedAwardsTotalCompensationCostNotYetRecognized"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_10c03073-b5ce-4a89-885d-4642fad84f63" xlink:to="loc_us-gaap_EmployeeServiceShareBasedCompensationNonvestedAwardsTotalCompensationCostNotYetRecognized_a1cda173-475b-49f6-a4fd-a03ac2d8f983" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_SharebasedCompensationArrangementNonemployeeWeightedAverageRemainingVestingPeriod_aad371d9-1743-41c6-b538-247f821588c5" xlink:href="trup-20201231.xsd#trup_SharebasedCompensationArrangementNonemployeeWeightedAverageRemainingVestingPeriod"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_10c03073-b5ce-4a89-885d-4642fad84f63" xlink:to="loc_trup_SharebasedCompensationArrangementNonemployeeWeightedAverageRemainingVestingPeriod_aad371d9-1743-41c6-b538-247f821588c5" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardOptionsOutstandingWeightedAverageRemainingContractualTerm2_852a2a51-27c6-42c1-a774-39a6395849b9" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardOptionsOutstandingWeightedAverageRemainingContractualTerm2"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_10c03073-b5ce-4a89-885d-4642fad84f63" xlink:to="loc_us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardOptionsOutstandingWeightedAverageRemainingContractualTerm2_852a2a51-27c6-42c1-a774-39a6395849b9" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable_4c11555f-d134-478f-a1ca-be5136652352" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_10c03073-b5ce-4a89-885d-4642fad84f63" xlink:to="loc_us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable_4c11555f-d134-478f-a1ca-be5136652352" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementClassOfStockAxis_b456ba1c-5be0-4147-82ea-347393765a68" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementClassOfStockAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable_4c11555f-d134-478f-a1ca-be5136652352" xlink:to="loc_us-gaap_StatementClassOfStockAxis_b456ba1c-5be0-4147-82ea-347393765a68" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_b456ba1c-5be0-4147-82ea-347393765a68_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementClassOfStockAxis_b456ba1c-5be0-4147-82ea-347393765a68" xlink:to="loc_us-gaap_ClassOfStockDomain_b456ba1c-5be0-4147-82ea-347393765a68_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_0f258a67-1a64-4281-b46b-8b46ef2e9f13" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementClassOfStockAxis_b456ba1c-5be0-4147-82ea-347393765a68" xlink:to="loc_us-gaap_ClassOfStockDomain_0f258a67-1a64-4281-b46b-8b46ef2e9f13" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_EmployeeStockOptionMember_442fcdfc-135b-4cff-bc47-334ff7f1f6f8" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_EmployeeStockOptionMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_0f258a67-1a64-4281-b46b-8b46ef2e9f13" xlink:to="loc_us-gaap_EmployeeStockOptionMember_442fcdfc-135b-4cff-bc47-334ff7f1f6f8" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_RestrictedStockUnitsRSUMember_5a749901-cfac-4419-af45-6aa220d20a05" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_RestrictedStockUnitsRSUMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_0f258a67-1a64-4281-b46b-8b46ef2e9f13" xlink:to="loc_us-gaap_RestrictedStockUnitsRSUMember_5a749901-cfac-4419-af45-6aa220d20a05" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/StockbasedCompensationExpenseCategoryDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#StockbasedCompensationExpenseCategoryDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockbasedCompensationExpenseCategoryDetails" xlink:type="extended" id="ib1f0732ae98a4108adfe91c6454ad0f6_StockbasedCompensationExpenseCategoryDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_EmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsLineItems_a69e7323-d655-437c-8a4e-dc58ab5fa388" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_EmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Restrictedstockoutstanding_d99b5c01-25f7-44a2-bc11-9e119d8fed33" xlink:href="trup-20201231.xsd#trup_Restrictedstockoutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsLineItems_a69e7323-d655-437c-8a4e-dc58ab5fa388" xlink:to="loc_trup_Restrictedstockoutstanding_d99b5c01-25f7-44a2-bc11-9e119d8fed33" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardCompensationCost1_2f430817-e534-49a7-aeb2-82133ef5dd33" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardCompensationCost1"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsLineItems_a69e7323-d655-437c-8a4e-dc58ab5fa388" xlink:to="loc_us-gaap_SharebasedCompensationArrangementBySharebasedPaymentAwardCompensationCost1_2f430817-e534-49a7-aeb2-82133ef5dd33" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_SharebasedCompensationArrangementbySharebasedPaymentAwardCapitalizedCost_34a72707-9b67-4ee8-b74d-e8c115c412cc" xlink:href="trup-20201231.xsd#trup_SharebasedCompensationArrangementbySharebasedPaymentAwardCapitalizedCost"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsLineItems_a69e7323-d655-437c-8a4e-dc58ab5fa388" xlink:to="loc_trup_SharebasedCompensationArrangementbySharebasedPaymentAwardCapitalizedCost_34a72707-9b67-4ee8-b74d-e8c115c412cc" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_EmployeeServiceShareBasedCompensationNonvestedAwardsTotalCompensationCostNotYetRecognized_aeff1349-6b18-4cc8-9f43-ea862f164cbc" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_EmployeeServiceShareBasedCompensationNonvestedAwardsTotalCompensationCostNotYetRecognized"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsLineItems_a69e7323-d655-437c-8a4e-dc58ab5fa388" xlink:to="loc_us-gaap_EmployeeServiceShareBasedCompensationNonvestedAwardsTotalCompensationCostNotYetRecognized_aeff1349-6b18-4cc8-9f43-ea862f164cbc" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_SharebasedCompensationArrangementNonemployeeWeightedAverageRemainingVestingPeriod_0234da37-b226-4ccb-b099-bdcae86e67cc" xlink:href="trup-20201231.xsd#trup_SharebasedCompensationArrangementNonemployeeWeightedAverageRemainingVestingPeriod"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsLineItems_a69e7323-d655-437c-8a4e-dc58ab5fa388" xlink:to="loc_trup_SharebasedCompensationArrangementNonemployeeWeightedAverageRemainingVestingPeriod_0234da37-b226-4ccb-b099-bdcae86e67cc" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AdjustmentsToAdditionalPaidInCapitalSharebasedCompensationRequisiteServicePeriodRecognitionValue_92dd3740-8b02-4a9a-ad33-d29d999ed847" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AdjustmentsToAdditionalPaidInCapitalSharebasedCompensationRequisiteServicePeriodRecognitionValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_EmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsLineItems_a69e7323-d655-437c-8a4e-dc58ab5fa388" xlink:to="loc_us-gaap_AdjustmentsToAdditionalPaidInCapitalSharebasedCompensationRequisiteServicePeriodRecognitionValue_92dd3740-8b02-4a9a-ad33-d29d999ed847" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ScheduleOfEmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsTable_c51ec68b-30ed-4e55-90e5-87da3787c2fd" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ScheduleOfEmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_EmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsLineItems_a69e7323-d655-437c-8a4e-dc58ab5fa388" xlink:to="loc_us-gaap_ScheduleOfEmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsTable_c51ec68b-30ed-4e55-90e5-87da3787c2fd" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementEquityComponentsAxis_ec348803-783c-48b4-8b02-394d8a5e6558" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementEquityComponentsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfEmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsTable_c51ec68b-30ed-4e55-90e5-87da3787c2fd" xlink:to="loc_us-gaap_StatementEquityComponentsAxis_ec348803-783c-48b4-8b02-394d8a5e6558" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_EquityComponentDomain_ec348803-783c-48b4-8b02-394d8a5e6558_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_EquityComponentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementEquityComponentsAxis_ec348803-783c-48b4-8b02-394d8a5e6558" xlink:to="loc_us-gaap_EquityComponentDomain_ec348803-783c-48b4-8b02-394d8a5e6558_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_EquityComponentDomain_8b4ded6c-f720-4323-bcc0-a16189495f43" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_EquityComponentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementEquityComponentsAxis_ec348803-783c-48b4-8b02-394d8a5e6558" xlink:to="loc_us-gaap_EquityComponentDomain_8b4ded6c-f720-4323-bcc0-a16189495f43" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementClassOfStockAxis_1cf00feb-37fa-4c1f-b034-d3b9a29be414" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementClassOfStockAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfEmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsTable_c51ec68b-30ed-4e55-90e5-87da3787c2fd" xlink:to="loc_us-gaap_StatementClassOfStockAxis_1cf00feb-37fa-4c1f-b034-d3b9a29be414" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_1cf00feb-37fa-4c1f-b034-d3b9a29be414_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementClassOfStockAxis_1cf00feb-37fa-4c1f-b034-d3b9a29be414" xlink:to="loc_us-gaap_ClassOfStockDomain_1cf00feb-37fa-4c1f-b034-d3b9a29be414_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_ad0b0849-ea40-4b7f-bbdf-3ceae47f6f98" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementClassOfStockAxis_1cf00feb-37fa-4c1f-b034-d3b9a29be414" xlink:to="loc_us-gaap_ClassOfStockDomain_ad0b0849-ea40-4b7f-bbdf-3ceae47f6f98" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_RestrictedStockUnitsRSUMember_57835757-1351-40f5-99e8-9d6d7c0692a8" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_RestrictedStockUnitsRSUMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_ad0b0849-ea40-4b7f-bbdf-3ceae47f6f98" xlink:to="loc_us-gaap_RestrictedStockUnitsRSUMember_57835757-1351-40f5-99e8-9d6d7c0692a8" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeStatementLocationAxis_c39c941b-efe4-4bcd-b589-1474e8698c82" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeStatementLocationAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfEmployeeServiceShareBasedCompensationAllocationOfRecognizedPeriodCostsTable_c51ec68b-30ed-4e55-90e5-87da3787c2fd" xlink:to="loc_us-gaap_IncomeStatementLocationAxis_c39c941b-efe4-4bcd-b589-1474e8698c82" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeStatementLocationDomain_c39c941b-efe4-4bcd-b589-1474e8698c82_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeStatementLocationDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_IncomeStatementLocationAxis_c39c941b-efe4-4bcd-b589-1474e8698c82" xlink:to="loc_us-gaap_IncomeStatementLocationDomain_c39c941b-efe4-4bcd-b589-1474e8698c82_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeStatementLocationDomain_46cff3b7-dc5d-4d02-ada3-2d51c81fcd38" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeStatementLocationDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_IncomeStatementLocationAxis_c39c941b-efe4-4bcd-b589-1474e8698c82" xlink:to="loc_us-gaap_IncomeStatementLocationDomain_46cff3b7-dc5d-4d02-ada3-2d51c81fcd38" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_ClaimsExpenseMember_8b807d25-d989-434a-8580-236f68bcbbbc" xlink:href="trup-20201231.xsd#trup_ClaimsExpenseMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncomeStatementLocationDomain_46cff3b7-dc5d-4d02-ada3-2d51c81fcd38" xlink:to="loc_trup_ClaimsExpenseMember_8b807d25-d989-434a-8580-236f68bcbbbc" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_OtherCostofSalesMember_e5ed04e5-51b1-4438-bc32-ec23347582e8" xlink:href="trup-20201231.xsd#trup_OtherCostofSalesMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncomeStatementLocationDomain_46cff3b7-dc5d-4d02-ada3-2d51c81fcd38" xlink:to="loc_trup_OtherCostofSalesMember_e5ed04e5-51b1-4438-bc32-ec23347582e8" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SellingAndMarketingExpenseMember_0f7cacbd-d508-4523-ae43-9b23e891e342" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SellingAndMarketingExpenseMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncomeStatementLocationDomain_46cff3b7-dc5d-4d02-ada3-2d51c81fcd38" xlink:to="loc_us-gaap_SellingAndMarketingExpenseMember_0f7cacbd-d508-4523-ae43-9b23e891e342" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ResearchAndDevelopmentExpenseMember_e060facc-6d5f-41cc-9aab-e26b27bb8336" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ResearchAndDevelopmentExpenseMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncomeStatementLocationDomain_46cff3b7-dc5d-4d02-ada3-2d51c81fcd38" xlink:to="loc_us-gaap_ResearchAndDevelopmentExpenseMember_e060facc-6d5f-41cc-9aab-e26b27bb8336" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_GeneralAndAdministrativeExpenseMember_6d99e8f9-3e8c-4408-a9d3-fe0e428b0ced" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_GeneralAndAdministrativeExpenseMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncomeStatementLocationDomain_46cff3b7-dc5d-4d02-ada3-2d51c81fcd38" xlink:to="loc_us-gaap_GeneralAndAdministrativeExpenseMember_6d99e8f9-3e8c-4408-a9d3-fe0e428b0ced" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_TotalExpenseMember_311c2060-ceeb-4699-abe3-b0e8f379a489" xlink:href="trup-20201231.xsd#trup_TotalExpenseMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_IncomeStatementLocationDomain_46cff3b7-dc5d-4d02-ada3-2d51c81fcd38" xlink:to="loc_trup_TotalExpenseMember_311c2060-ceeb-4699-abe3-b0e8f379a489" xlink:type="arc" order="5"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/StockbasedCompensationValuationAssumptionsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#StockbasedCompensationValuationAssumptionsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockbasedCompensationValuationAssumptionsDetails" xlink:type="extended" id="i74cd4fc1412044049e7b1d23bd262d12_StockbasedCompensationValuationAssumptionsDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/StockbasedCompensationOptionActivityDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#StockbasedCompensationOptionActivityDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockbasedCompensationOptionActivityDetails" xlink:type="extended" id="iad16b41d5a2c459da9323e2794bef7c9_StockbasedCompensationOptionActivityDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/StockbasedCompensationOptionsGrantedDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#StockbasedCompensationOptionsGrantedDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockbasedCompensationOptionsGrantedDetails" xlink:type="extended" id="i882dc65b10184132aa890b858453ec7c_StockbasedCompensationOptionsGrantedDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/StockbasedCompensationRestrictedStockAwardsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#StockbasedCompensationRestrictedStockAwardsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockbasedCompensationRestrictedStockAwardsDetails" xlink:type="extended" id="ief6397ce7e1b4ef7863db58071504d92_StockbasedCompensationRestrictedStockAwardsDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_d23aceb6-6275-47e5-887b-db00fa68f396" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedRollForward_8a58986d-7c28-4c85-aef7-630163e61f4a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedRollForward"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_d23aceb6-6275-47e5-887b-db00fa68f396" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedRollForward_8a58986d-7c28-4c85-aef7-630163e61f4a" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedNumber_408f0dd7-3386-418a-afd8-f3bd84abd3dc" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedNumber"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedRollForward_8a58986d-7c28-4c85-aef7-630163e61f4a" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedNumber_408f0dd7-3386-418a-afd8-f3bd84abd3dc" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsGrantsInPeriod_937b4d95-9cc0-498d-92f3-205557e59d8c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsGrantsInPeriod"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedRollForward_8a58986d-7c28-4c85-aef7-630163e61f4a" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsGrantsInPeriod_937b4d95-9cc0-498d-92f3-205557e59d8c" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsVestedInPeriod_ddb8625d-4ae9-43ea-852a-15b720c7ca03" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsVestedInPeriod"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedRollForward_8a58986d-7c28-4c85-aef7-630163e61f4a" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsVestedInPeriod_ddb8625d-4ae9-43ea-852a-15b720c7ca03" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsForfeitedInPeriod_865d22bb-b98b-40f0-97e3-9de8bf7ae79c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsForfeitedInPeriod"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedRollForward_8a58986d-7c28-4c85-aef7-630163e61f4a" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsForfeitedInPeriod_865d22bb-b98b-40f0-97e3-9de8bf7ae79c" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedNumber_6dc0fccb-5aa2-4b81-910b-641c4b28e8b0" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedNumber"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_SharebasedCompensationArrangementbySharebasedPaymentAwardEquityInstrumentsOtherthanOptionsWeightedAverageGrantDateFairValueRollForward_700e304e-39c7-4e26-8d11-be8a2385e5bc" xlink:href="trup-20201231.xsd#trup_SharebasedCompensationArrangementbySharebasedPaymentAwardEquityInstrumentsOtherthanOptionsWeightedAverageGrantDateFairValueRollForward"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_d23aceb6-6275-47e5-887b-db00fa68f396" xlink:to="loc_trup_SharebasedCompensationArrangementbySharebasedPaymentAwardEquityInstrumentsOtherthanOptionsWeightedAverageGrantDateFairValueRollForward_700e304e-39c7-4e26-8d11-be8a2385e5bc" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedWeightedAverageGrantDateFairValue_43a65d0d-eede-4d57-bd92-f9a2e8aeb2a4" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedWeightedAverageGrantDateFairValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_SharebasedCompensationArrangementbySharebasedPaymentAwardEquityInstrumentsOtherthanOptionsWeightedAverageGrantDateFairValueRollForward_700e304e-39c7-4e26-8d11-be8a2385e5bc" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedWeightedAverageGrantDateFairValue_43a65d0d-eede-4d57-bd92-f9a2e8aeb2a4" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsGrantsInPeriodWeightedAverageGrantDateFairValue_365288b6-8acd-4048-8501-9992631aea5b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsGrantsInPeriodWeightedAverageGrantDateFairValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_SharebasedCompensationArrangementbySharebasedPaymentAwardEquityInstrumentsOtherthanOptionsWeightedAverageGrantDateFairValueRollForward_700e304e-39c7-4e26-8d11-be8a2385e5bc" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsGrantsInPeriodWeightedAverageGrantDateFairValue_365288b6-8acd-4048-8501-9992631aea5b" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsVestedInPeriodWeightedAverageGrantDateFairValue_04abd608-036f-4497-a7a3-7014c5e447cf" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsVestedInPeriodWeightedAverageGrantDateFairValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_SharebasedCompensationArrangementbySharebasedPaymentAwardEquityInstrumentsOtherthanOptionsWeightedAverageGrantDateFairValueRollForward_700e304e-39c7-4e26-8d11-be8a2385e5bc" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsVestedInPeriodWeightedAverageGrantDateFairValue_04abd608-036f-4497-a7a3-7014c5e447cf" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsForfeituresWeightedAverageGrantDateFairValue_4697b2c3-41e9-453e-a31d-06aa19523bf7" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsForfeituresWeightedAverageGrantDateFairValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_trup_SharebasedCompensationArrangementbySharebasedPaymentAwardEquityInstrumentsOtherthanOptionsWeightedAverageGrantDateFairValueRollForward_700e304e-39c7-4e26-8d11-be8a2385e5bc" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsForfeituresWeightedAverageGrantDateFairValue_4697b2c3-41e9-453e-a31d-06aa19523bf7" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedWeightedAverageGrantDateFairValue_2eb51c4a-f144-40d4-8023-f98dab1e3595" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsNonvestedWeightedAverageGrantDateFairValue"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable_1b44c99c-ff16-4d13-b19a-98fc57a01066" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems_d23aceb6-6275-47e5-887b-db00fa68f396" xlink:to="loc_us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable_1b44c99c-ff16-4d13-b19a-98fc57a01066" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AwardTypeAxis_eea514db-a75e-48b5-a626-610242a5560c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AwardTypeAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable_1b44c99c-ff16-4d13-b19a-98fc57a01066" xlink:to="loc_us-gaap_AwardTypeAxis_eea514db-a75e-48b5-a626-610242a5560c" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_eea514db-a75e-48b5-a626-610242a5560c_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_AwardTypeAxis_eea514db-a75e-48b5-a626-610242a5560c" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_eea514db-a75e-48b5-a626-610242a5560c_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_ca97388d-4a63-4d33-bd54-7b8e58238e73" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_AwardTypeAxis_eea514db-a75e-48b5-a626-610242a5560c" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_ca97388d-4a63-4d33-bd54-7b8e58238e73" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_RestrictedStockMember_6f4940ba-3214-4dad-ade6-fcda781e0622" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_RestrictedStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_ca97388d-4a63-4d33-bd54-7b8e58238e73" xlink:to="loc_us-gaap_RestrictedStockMember_6f4940ba-3214-4dad-ade6-fcda781e0622" xlink:type="arc" order="0"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/LeasesNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#LeasesNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/LeasesNotes" xlink:type="extended" id="ia4bd11561f8a4090b88d88734745494c_LeasesNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/LeasesTables" xlink:type="simple" xlink:href="trup-20201231.xsd#LeasesTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/LeasesTables" xlink:type="extended" id="ie58089efdbcf48c98c0d6174ebbb9cc4_LeasesTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/LeasesDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#LeasesDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/LeasesDetails" xlink:type="extended" id="iacafcfd010d1495fbc305dc325fcd599_LeasesDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/StockholdersEquityNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#StockholdersEquityNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockholdersEquityNotes" xlink:type="extended" id="i3465009174414570a6471b64db0ac43e_StockholdersEquityNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/StockholdersEquityNarrativeDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#StockholdersEquityNarrativeDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/StockholdersEquityNarrativeDetails" xlink:type="extended" id="ib5e255684f734482a8ddff616db6b1c1_StockholdersEquityNarrativeDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/Segments" xlink:type="simple" xlink:href="trup-20201231.xsd#Segments"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/Segments" xlink:type="extended" id="ie057483b147246b4a9db0f33bf25372a_Segments"/>
<link:roleRef roleURI="http://www.trupanion.com/role/SegmentsTables" xlink:type="simple" xlink:href="trup-20201231.xsd#SegmentsTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/SegmentsTables" xlink:type="extended" id="ie13b0b3e5944492094e73b26c195939c_SegmentsTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/SegmentsDetailsBusinessSegment" xlink:type="simple" xlink:href="trup-20201231.xsd#SegmentsDetailsBusinessSegment"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/SegmentsDetailsBusinessSegment" xlink:type="extended" id="i5f6956bb7fe847af885cfa0feba43a57_SegmentsDetailsBusinessSegment">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentReportingInformationLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_Revenues_03c86681-eeef-49e5-93fc-edb7894a61b3" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_Revenues"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_us-gaap_Revenues_03c86681-eeef-49e5-93fc-edb7894a61b3" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_ClaimsExpense_c5326056-bfce-4847-99bb-b1d0196590e9" xlink:href="trup-20201231.xsd#trup_ClaimsExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_trup_ClaimsExpense_c5326056-bfce-4847-99bb-b1d0196590e9" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_OtherCostofServicesSold_c1f80014-86fd-49b6-b1c5-ecaacf63f9ea" xlink:href="trup-20201231.xsd#trup_OtherCostofServicesSold"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_trup_OtherCostofServicesSold_c1f80014-86fd-49b6-b1c5-ecaacf63f9ea" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_TechnologyandDevelopmentExpense_d22f7825-9d89-4baf-b90d-20a97ea4e643" xlink:href="trup-20201231.xsd#trup_TechnologyandDevelopmentExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_trup_TechnologyandDevelopmentExpense_d22f7825-9d89-4baf-b90d-20a97ea4e643" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_GeneralAndAdministrativeExpense_d377e76e-cff2-4ad0-93f7-fd054acb5dc2" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_GeneralAndAdministrativeExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_us-gaap_GeneralAndAdministrativeExpense_d377e76e-cff2-4ad0-93f7-fd054acb5dc2" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SellingAndMarketingExpense_b792662d-c8b2-45a3-8071-d71ce48affce" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SellingAndMarketingExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_us-gaap_SellingAndMarketingExpense_b792662d-c8b2-45a3-8071-d71ce48affce" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CostOfGoodsAndServicesSoldDepreciationAndAmortization_1cebe919-8eb0-43f1-9434-65277062ee67" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CostOfGoodsAndServicesSoldDepreciationAndAmortization"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_us-gaap_CostOfGoodsAndServicesSoldDepreciationAndAmortization_1cebe919-8eb0-43f1-9434-65277062ee67" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeLossFromEquityMethodInvestments_4adf0bb0-b604-481e-b618-9c4743a3bd68" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeLossFromEquityMethodInvestments"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_us-gaap_IncomeLossFromEquityMethodInvestments_4adf0bb0-b604-481e-b618-9c4743a3bd68" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingIncomeLoss_e83f17dc-410d-4c06-903d-2c63b55a4350" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OperatingIncomeLoss"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_us-gaap_OperatingIncomeLoss_e83f17dc-410d-4c06-903d-2c63b55a4350" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ScheduleOfSegmentReportingInformationBySegmentTable_2ef74f9c-c68c-4a6a-becd-8a53a9a56c66" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ScheduleOfSegmentReportingInformationBySegmentTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_1cf6128b-4205-4541-a94d-aed031052cb1" xlink:to="loc_us-gaap_ScheduleOfSegmentReportingInformationBySegmentTable_2ef74f9c-c68c-4a6a-becd-8a53a9a56c66" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementBusinessSegmentsAxis_5bf54222-9a75-44af-a3b0-2656ed84b9cf" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementBusinessSegmentsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfSegmentReportingInformationBySegmentTable_2ef74f9c-c68c-4a6a-becd-8a53a9a56c66" xlink:to="loc_us-gaap_StatementBusinessSegmentsAxis_5bf54222-9a75-44af-a3b0-2656ed84b9cf" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_5bf54222-9a75-44af-a3b0-2656ed84b9cf_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_5bf54222-9a75-44af-a3b0-2656ed84b9cf" xlink:to="loc_us-gaap_SegmentDomain_5bf54222-9a75-44af-a3b0-2656ed84b9cf_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_2189cc4c-a377-4f36-bd87-ab129239c2a1" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_5bf54222-9a75-44af-a3b0-2656ed84b9cf" xlink:to="loc_us-gaap_SegmentDomain_2189cc4c-a377-4f36-bd87-ab129239c2a1" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_SubscriptionbusinessMember_76363d56-7e93-440f-bdcc-aa8e0a91bbd1" xlink:href="trup-20201231.xsd#trup_SubscriptionbusinessMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_2189cc4c-a377-4f36-bd87-ab129239c2a1" xlink:to="loc_trup_SubscriptionbusinessMember_76363d56-7e93-440f-bdcc-aa8e0a91bbd1" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AllOtherSegmentsMember_132a02ba-436d-4e84-8b7b-d39b8f5ef011" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AllOtherSegmentsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_2189cc4c-a377-4f36-bd87-ab129239c2a1" xlink:to="loc_us-gaap_AllOtherSegmentsMember_132a02ba-436d-4e84-8b7b-d39b8f5ef011" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/SegmentsDetailsRevenuebyGeography" xlink:type="simple" xlink:href="trup-20201231.xsd#SegmentsDetailsRevenuebyGeography"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/SegmentsDetailsRevenuebyGeography" xlink:type="extended" id="i6f8f17a266994e9c9f1e29ef20dce3ba_SegmentsDetailsRevenuebyGeography">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentReportingInformationLineItems_0357dc9d-5652-44c1-b93e-508178466057" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentReportingInformationLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_Revenues_05ce04c6-17e1-4637-9564-81da80295003" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_Revenues"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_0357dc9d-5652-44c1-b93e-508178466057" xlink:to="loc_us-gaap_Revenues_05ce04c6-17e1-4637-9564-81da80295003" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ScheduleOfSegmentReportingInformationBySegmentTable_ea6c2fc1-af01-497e-824d-7af2259412d5" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ScheduleOfSegmentReportingInformationBySegmentTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_SegmentReportingInformationLineItems_0357dc9d-5652-44c1-b93e-508178466057" xlink:to="loc_us-gaap_ScheduleOfSegmentReportingInformationBySegmentTable_ea6c2fc1-af01-497e-824d-7af2259412d5" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_StatementGeographicalAxis_3a7f24ef-499d-4365-82b4-050e281eceb5" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_StatementGeographicalAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_ScheduleOfSegmentReportingInformationBySegmentTable_ea6c2fc1-af01-497e-824d-7af2259412d5" xlink:to="loc_srt_StatementGeographicalAxis_3a7f24ef-499d-4365-82b4-050e281eceb5" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_SegmentGeographicalDomain_3a7f24ef-499d-4365-82b4-050e281eceb5_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_SegmentGeographicalDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_StatementGeographicalAxis_3a7f24ef-499d-4365-82b4-050e281eceb5" xlink:to="loc_srt_SegmentGeographicalDomain_3a7f24ef-499d-4365-82b4-050e281eceb5_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_SegmentGeographicalDomain_b321ae73-db50-41ba-a13c-71d39f916948" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_SegmentGeographicalDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_StatementGeographicalAxis_3a7f24ef-499d-4365-82b4-050e281eceb5" xlink:to="loc_srt_SegmentGeographicalDomain_b321ae73-db50-41ba-a13c-71d39f916948" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_country_US_826126e2-0151-495d-bf23-a7859f11a8de" xlink:href="https://xbrl.sec.gov/country/2020/country-2020-01-31.xsd#country_US"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_SegmentGeographicalDomain_b321ae73-db50-41ba-a13c-71d39f916948" xlink:to="loc_country_US_826126e2-0151-495d-bf23-a7859f11a8de" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_country_CA_34d0ca36-12bc-4e64-948b-924699b0faae" xlink:href="https://xbrl.sec.gov/country/2020/country-2020-01-31.xsd#country_CA"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_SegmentGeographicalDomain_b321ae73-db50-41ba-a13c-71d39f916948" xlink:to="loc_country_CA_34d0ca36-12bc-4e64-948b-924699b0faae" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/DividendRestrictionsStatutorySurplusNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#DividendRestrictionsStatutorySurplusNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/DividendRestrictionsStatutorySurplusNotes" xlink:type="extended" id="i29f6c9679589444cb6a383f635e532db_DividendRestrictionsStatutorySurplusNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/DividendRestrictionsStatutorySurplusTables" xlink:type="simple" xlink:href="trup-20201231.xsd#DividendRestrictionsStatutorySurplusTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/DividendRestrictionsStatutorySurplusTables" xlink:type="extended" id="i4512a60793354a18847b38e735235a67_DividendRestrictionsStatutorySurplusTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/DividendRestrictionsStatutorySurplusDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#DividendRestrictionsStatutorySurplusDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/DividendRestrictionsStatutorySurplusDetails" xlink:type="extended" id="i3d86769b257f4661ad615ac730242bc7_DividendRestrictionsStatutorySurplusDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IncomeTaxesNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#IncomeTaxesNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IncomeTaxesNotes" xlink:type="extended" id="i43e1a702991b46c490faef92d75a43a3_IncomeTaxesNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IncomeTaxesTables" xlink:type="simple" xlink:href="trup-20201231.xsd#IncomeTaxesTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IncomeTaxesTables" xlink:type="extended" id="ia8d9b06181f84325842f651a7086e2c5_IncomeTaxesTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IncomeTaxesIncomebeforetaxesDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#IncomeTaxesIncomebeforetaxesDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IncomeTaxesIncomebeforetaxesDetails" xlink:type="extended" id="i21cb752f675a4d5d98bbb02dafb23f9c_IncomeTaxesIncomebeforetaxesDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IncomeTaxesIncometaxbenefitsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#IncomeTaxesIncometaxbenefitsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IncomeTaxesIncometaxbenefitsDetails" xlink:type="extended" id="id41b3624df3740d7bab0cc73ea7bdb12_IncomeTaxesIncometaxbenefitsDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IncomeTaxesTaxRateReconciliationDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#IncomeTaxesTaxRateReconciliationDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IncomeTaxesTaxRateReconciliationDetails" xlink:type="extended" id="i33643731d3e444a0b3a477a084509828_IncomeTaxesTaxRateReconciliationDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IncomeTaxesDeferredtaxassetsandliabilitiesDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#IncomeTaxesDeferredtaxassetsandliabilitiesDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IncomeTaxesDeferredtaxassetsandliabilitiesDetails" xlink:type="extended" id="if291e37e9f5a4875975441bf493c0182_IncomeTaxesDeferredtaxassetsandliabilitiesDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/IncomeTaxesUnrecognizedTaxBenefitsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#IncomeTaxesUnrecognizedTaxBenefitsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/IncomeTaxesUnrecognizedTaxBenefitsDetails" xlink:type="extended" id="ie6fe6c67d4e745a08ae191f9fc57def3_IncomeTaxesUnrecognizedTaxBenefitsDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/RetirementPlanNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#RetirementPlanNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/RetirementPlanNotes" xlink:type="extended" id="if5d2c0c866d84bc3a3f294e9cb423692_RetirementPlanNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/RetirementPlanDetailsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#RetirementPlanDetailsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/RetirementPlanDetailsDetails" xlink:type="extended" id="i5b67a10111224919839ba09e7510435d_RetirementPlanDetailsDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/RelatedPartyDisclosures" xlink:type="simple" xlink:href="trup-20201231.xsd#RelatedPartyDisclosures"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/RelatedPartyDisclosures" xlink:type="extended" id="if0cf68ff6e4d421b9506aa5177f35673_RelatedPartyDisclosures"/>
<link:roleRef roleURI="http://www.trupanion.com/role/RelatedPartyDisclosuresTables" xlink:type="simple" xlink:href="trup-20201231.xsd#RelatedPartyDisclosuresTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/RelatedPartyDisclosuresTables" xlink:type="extended" id="ie6af24a65aa5481a979acd682db143f6_RelatedPartyDisclosuresTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/RelatedPartyDisclosuresDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#RelatedPartyDisclosuresDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/RelatedPartyDisclosuresDetails" xlink:type="extended" id="i5352481845ce44ebaf86d4f8ff60c9fb_RelatedPartyDisclosuresDetails"/>
<link:roleRef roleURI="http://www.trupanion.com/role/QuarterlyFinancialInformationNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#QuarterlyFinancialInformationNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/QuarterlyFinancialInformationNotes" xlink:type="extended" id="i34cf783b54e947c18c1c239ce0b1a915_QuarterlyFinancialInformationNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/QuarterlyFinancialInformationTables" xlink:type="simple" xlink:href="trup-20201231.xsd#QuarterlyFinancialInformationTables"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/QuarterlyFinancialInformationTables" xlink:type="extended" id="icc65a1fa6fc249c5ba961285308eb128_QuarterlyFinancialInformationTables"/>
<link:roleRef roleURI="http://www.trupanion.com/role/QuarterlyFinancialInformationDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#QuarterlyFinancialInformationDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/QuarterlyFinancialInformationDetails" xlink:type="extended" id="i13dccad5e24d4aa180be3a6c594ceeb9_QuarterlyFinancialInformationDetails">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementLineItems_992be43d-3c41-480e-97e6-2d3181a592de" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementTable_2a68dc0f-c867-413c-b305-83e042092c4a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_StatementLineItems_992be43d-3c41-480e-97e6-2d3181a592de" xlink:to="loc_us-gaap_StatementTable_2a68dc0f-c867-413c-b305-83e042092c4a" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementBusinessSegmentsAxis_c0ab112b-9319-4f6f-b8de-1a7a2bf09255" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementBusinessSegmentsAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_StatementTable_2a68dc0f-c867-413c-b305-83e042092c4a" xlink:to="loc_us-gaap_StatementBusinessSegmentsAxis_c0ab112b-9319-4f6f-b8de-1a7a2bf09255" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_c0ab112b-9319-4f6f-b8de-1a7a2bf09255_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_c0ab112b-9319-4f6f-b8de-1a7a2bf09255" xlink:to="loc_us-gaap_SegmentDomain_c0ab112b-9319-4f6f-b8de-1a7a2bf09255_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SegmentDomain_519314d0-bdf7-4e4d-ad5a-25af35bb30b9" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SegmentDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementBusinessSegmentsAxis_c0ab112b-9319-4f6f-b8de-1a7a2bf09255" xlink:to="loc_us-gaap_SegmentDomain_519314d0-bdf7-4e4d-ad5a-25af35bb30b9" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AllOtherSegmentsMember_f69376fe-a4fd-4c97-a469-2eec437f77d6" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AllOtherSegmentsMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_519314d0-bdf7-4e4d-ad5a-25af35bb30b9" xlink:to="loc_us-gaap_AllOtherSegmentsMember_f69376fe-a4fd-4c97-a469-2eec437f77d6" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_SubscriptionbusinessMember_e79ea07c-ff22-48a7-98b0-7efb1621db42" xlink:href="trup-20201231.xsd#trup_SubscriptionbusinessMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_SegmentDomain_519314d0-bdf7-4e4d-ad5a-25af35bb30b9" xlink:to="loc_trup_SubscriptionbusinessMember_e79ea07c-ff22-48a7-98b0-7efb1621db42" xlink:type="arc" order="1"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/Schedule1ParentOnlyDisclosuresScheduleNotes" xlink:type="simple" xlink:href="trup-20201231.xsd#Schedule1ParentOnlyDisclosuresScheduleNotes"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/Schedule1ParentOnlyDisclosuresScheduleNotes" xlink:type="extended" id="ibfdf30a746664d29802ae2abb7023260_Schedule1ParentOnlyDisclosuresScheduleNotes"/>
<link:roleRef roleURI="http://www.trupanion.com/role/Schedule1ParentOnlyDisclosuresScheduleCondensedStatementofComprehensiveLossDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#Schedule1ParentOnlyDisclosuresScheduleCondensedStatementofComprehensiveLossDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/Schedule1ParentOnlyDisclosuresScheduleCondensedStatementofComprehensiveLossDetails" xlink:type="extended" id="i51136bcdbccd46e1ae7d853fb80b32b7_Schedule1ParentOnlyDisclosuresScheduleCondensedStatementofComprehensiveLossDetails">
<link:loc xlink:type="locator" xlink:label="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_CondensedFinancialStatementsCaptionsLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_Veterinaryinvoiceexpense_a8320260-91fd-4f80-a7dd-163d93976c77" xlink:href="trup-20201231.xsd#trup_Veterinaryinvoiceexpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_trup_Veterinaryinvoiceexpense_a8320260-91fd-4f80-a7dd-163d93976c77" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherCostOfOperatingRevenue_d66788be-800d-4c84-aa8d-1485e7cda009" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherCostOfOperatingRevenue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_OtherCostOfOperatingRevenue_d66788be-800d-4c84-aa8d-1485e7cda009" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_TechnologyandDevelopmentExpense_1dc231a4-d2fa-4a07-811e-22c593c8efc8" xlink:href="trup-20201231.xsd#trup_TechnologyandDevelopmentExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_trup_TechnologyandDevelopmentExpense_1dc231a4-d2fa-4a07-811e-22c593c8efc8" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_GeneralAndAdministrativeExpense_5946057d-8ad7-4429-85a1-4bfdc6f4be8e" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_GeneralAndAdministrativeExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_GeneralAndAdministrativeExpense_5946057d-8ad7-4429-85a1-4bfdc6f4be8e" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_SellingAndMarketingExpense_b6c942cf-8289-4f7c-a122-d18fd1ce120f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_SellingAndMarketingExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_SellingAndMarketingExpense_b6c942cf-8289-4f7c-a122-d18fd1ce120f" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CostOfGoodsAndServicesSoldDepreciationAndAmortization_1350ac81-744b-4a36-8201-5bfb0debada5" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CostOfGoodsAndServicesSoldDepreciationAndAmortization"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_CostOfGoodsAndServicesSoldDepreciationAndAmortization_1350ac81-744b-4a36-8201-5bfb0debada5" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CostsAndExpenses_b051b7a2-7fee-4332-b3c0-ac07c696b192" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CostsAndExpenses"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_CostsAndExpenses_b051b7a2-7fee-4332-b3c0-ac07c696b192" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeLossFromEquityMethodInvestments_749831e1-f0db-450b-9025-fc14ed5f5edc" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeLossFromEquityMethodInvestments"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_IncomeLossFromEquityMethodInvestments_749831e1-f0db-450b-9025-fc14ed5f5edc" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingIncomeLoss_9924a2ef-091e-4fa1-963a-6e7bb88db400" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OperatingIncomeLoss"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_OperatingIncomeLoss_9924a2ef-091e-4fa1-963a-6e7bb88db400" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_InterestExpense_ec12532a-af3d-487c-b716-03e39acda5b8" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_InterestExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_InterestExpense_ec12532a-af3d-487c-b716-03e39acda5b8" xlink:type="arc" order="9"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherNonoperatingIncomeExpense_1db0ab1d-84cc-46bc-99e3-a2073f2e8741" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherNonoperatingIncomeExpense"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_OtherNonoperatingIncomeExpense_1db0ab1d-84cc-46bc-99e3-a2073f2e8741" xlink:type="arc" order="10"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_LossBeforeEquityinUndistributedEarningsofSubsidiaries_1a6c26a0-dda7-4d17-b92c-ce821c0d3b0c" xlink:href="trup-20201231.xsd#trup_LossBeforeEquityinUndistributedEarningsofSubsidiaries"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_trup_LossBeforeEquityinUndistributedEarningsofSubsidiaries_1a6c26a0-dda7-4d17-b92c-ce821c0d3b0c" xlink:type="arc" order="11"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_IncomeTaxBenefitExpenseAttributabletoParent_ffa76093-8f69-457a-b713-fa42e0d667c6" xlink:href="trup-20201231.xsd#trup_IncomeTaxBenefitExpenseAttributabletoParent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_trup_IncomeTaxBenefitExpenseAttributabletoParent_ffa76093-8f69-457a-b713-fa42e0d667c6" xlink:type="arc" order="12"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_EquityLossInUndistributedEarningsOfSubsidiaries_c3aaeb8f-066b-4a50-aae0-26137ac92409" xlink:href="trup-20201231.xsd#trup_EquityLossInUndistributedEarningsOfSubsidiaries"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_trup_EquityLossInUndistributedEarningsOfSubsidiaries_c3aaeb8f-066b-4a50-aae0-26137ac92409" xlink:type="arc" order="13"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetIncomeLoss_cf3d5747-5e93-4061-838a-780ae9fc3fb5" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_NetIncomeLoss"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_NetIncomeLoss_cf3d5747-5e93-4061-838a-780ae9fc3fb5" xlink:type="arc" order="14"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent_128f57b7-3ea1-4c2e-adf5-30f1fb6f869a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent_128f57b7-3ea1-4c2e-adf5-30f1fb6f869a" xlink:type="arc" order="15"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ComprehensiveIncomeNetOfTax_73e9b481-cda5-4f01-93f4-53f377d9da96" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ComprehensiveIncomeNetOfTax"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_us-gaap_ComprehensiveIncomeNetOfTax_73e9b481-cda5-4f01-93f4-53f377d9da96" xlink:type="arc" order="16"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ScheduleOfCondensedFinancialStatementsTable_1ef0adfe-c735-46a9-8e54-a3de15b612cc" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ScheduleOfCondensedFinancialStatementsTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_7bac5119-5ac8-4da1-bf73-012f6d99947f" xlink:to="loc_srt_ScheduleOfCondensedFinancialStatementsTable_1ef0adfe-c735-46a9-8e54-a3de15b612cc" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesAxis_a9b6b1b3-7867-403d-90b1-7cb9f91a0cf3" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_srt_ScheduleOfCondensedFinancialStatementsTable_1ef0adfe-c735-46a9-8e54-a3de15b612cc" xlink:to="loc_srt_ConsolidatedEntitiesAxis_a9b6b1b3-7867-403d-90b1-7cb9f91a0cf3" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesDomain_a9b6b1b3-7867-403d-90b1-7cb9f91a0cf3_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_ConsolidatedEntitiesAxis_a9b6b1b3-7867-403d-90b1-7cb9f91a0cf3" xlink:to="loc_srt_ConsolidatedEntitiesDomain_a9b6b1b3-7867-403d-90b1-7cb9f91a0cf3_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesDomain_fb9307bf-f004-4561-916b-98bac80f03f0" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_ConsolidatedEntitiesAxis_a9b6b1b3-7867-403d-90b1-7cb9f91a0cf3" xlink:to="loc_srt_ConsolidatedEntitiesDomain_fb9307bf-f004-4561-916b-98bac80f03f0" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ParentCompanyMember_75dc5ba2-c4f6-48c7-8038-87d3312d2c25" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ParentCompanyMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_ConsolidatedEntitiesDomain_fb9307bf-f004-4561-916b-98bac80f03f0" xlink:to="loc_srt_ParentCompanyMember_75dc5ba2-c4f6-48c7-8038-87d3312d2c25" xlink:type="arc" order="0"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/Schedule1ParentOnlyDisclosuresScheduleCondensedBalanceSheetDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#Schedule1ParentOnlyDisclosuresScheduleCondensedBalanceSheetDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/Schedule1ParentOnlyDisclosuresScheduleCondensedBalanceSheetDetails" xlink:type="extended" id="ieff23498ecda48269dc9239ef79220bf_Schedule1ParentOnlyDisclosuresScheduleCondensedBalanceSheetDetails">
<link:loc xlink:type="locator" xlink:label="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_CondensedFinancialStatementsCaptionsLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CashEquivalentsAtCarryingValue_234ab0b7-20b7-48fc-9280-4e20908e3244" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CashEquivalentsAtCarryingValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_CashEquivalentsAtCarryingValue_234ab0b7-20b7-48fc-9280-4e20908e3244" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccountsAndOtherReceivablesNetCurrent_fda8de34-92c3-40ae-afd5-9639261d432f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AccountsAndOtherReceivablesNetCurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_AccountsAndOtherReceivablesNetCurrent_fda8de34-92c3-40ae-afd5-9639261d432f" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PrepaidExpenseAndOtherAssets_9cedaa7d-0fcc-4e8e-b036-708db180e5bd" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PrepaidExpenseAndOtherAssets"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_PrepaidExpenseAndOtherAssets_9cedaa7d-0fcc-4e8e-b036-708db180e5bd" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AssetsCurrent_bbc8c458-a4fd-4926-9b77-c129357a915b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AssetsCurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_AssetsCurrent_bbc8c458-a4fd-4926-9b77-c129357a915b" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_RestrictedCashAndCashEquivalents_1eccba65-2284-48da-bd74-09cde0363fae" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_RestrictedCashAndCashEquivalents"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_RestrictedCashAndCashEquivalents_1eccba65-2284-48da-bd74-09cde0363fae" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentNet_e8042a35-02a8-4df3-b788-6a4e8126267f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PropertyPlantAndEquipmentNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentNet_e8042a35-02a8-4df3-b788-6a4e8126267f" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IntangibleAssetsNetExcludingGoodwill_2ef3803e-168e-461d-80e8-beefbf362273" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IntangibleAssetsNetExcludingGoodwill"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_IntangibleAssetsNetExcludingGoodwill_2ef3803e-168e-461d-80e8-beefbf362273" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherAssetsNoncurrent_0ab76c3a-7869-44e0-9fcf-f8b31bd73b64" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherAssetsNoncurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_OtherAssetsNoncurrent_0ab76c3a-7869-44e0-9fcf-f8b31bd73b64" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_InvestmentsInAffiliatesSubsidiariesAssociatesAndJointVentures_4c9d7f21-c394-4b29-ab58-9dd0eaee7a63" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_InvestmentsInAffiliatesSubsidiariesAssociatesAndJointVentures"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_InvestmentsInAffiliatesSubsidiariesAssociatesAndJointVentures_4c9d7f21-c394-4b29-ab58-9dd0eaee7a63" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_Assets_5c051c91-9b1d-4549-b182-0c5bc764f829" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_Assets"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_Assets_5c051c91-9b1d-4549-b182-0c5bc764f829" xlink:type="arc" order="9"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccountsPayableAndAccruedLiabilitiesCurrent_3963f45f-a20e-4ae5-b75a-50cc676fe884" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AccountsPayableAndAccruedLiabilitiesCurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_AccountsPayableAndAccruedLiabilitiesCurrent_3963f45f-a20e-4ae5-b75a-50cc676fe884" xlink:type="arc" order="10"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccruedLiabilitiesCurrent_5506647c-7a8f-4314-bf0f-0df86b7f6403" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AccruedLiabilitiesCurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_AccruedLiabilitiesCurrent_5506647c-7a8f-4314-bf0f-0df86b7f6403" xlink:type="arc" order="11"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilitiesCurrent_9a4c66c6-676e-4de7-9f5e-e2641eaac0c0" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilitiesCurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_LiabilitiesCurrent_9a4c66c6-676e-4de7-9f5e-e2641eaac0c0" xlink:type="arc" order="12"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LongTermDebtNoncurrent_35923d6c-7a86-447b-9e0b-09431ad7e62b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LongTermDebtNoncurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_LongTermDebtNoncurrent_35923d6c-7a86-447b-9e0b-09431ad7e62b" xlink:type="arc" order="13"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_DeferredTaxAndOtherLiabilitiesNoncurrent_62dd4f8c-9bcf-4b2b-8054-7dd4936b36e5" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_DeferredTaxAndOtherLiabilitiesNoncurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_DeferredTaxAndOtherLiabilitiesNoncurrent_62dd4f8c-9bcf-4b2b-8054-7dd4936b36e5" xlink:type="arc" order="14"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherLiabilitiesNoncurrent_fde211a9-7a76-4d22-8517-1c0f871339fd" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherLiabilitiesNoncurrent"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_OtherLiabilitiesNoncurrent_fde211a9-7a76-4d22-8517-1c0f871339fd" xlink:type="arc" order="15"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_Liabilities_10da5223-2f6e-414b-8938-b81970b21be2" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_Liabilities"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_Liabilities_10da5223-2f6e-414b-8938-b81970b21be2" xlink:type="arc" order="16"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockValueOutstanding_3ad9fdcc-e6af-411d-b8ac-f325c266a56b" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockValueOutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_CommonStockValueOutstanding_3ad9fdcc-e6af-411d-b8ac-f325c266a56b" xlink:type="arc" order="17"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockValueOutstanding_bb2ea530-9973-4d25-a049-d8284405661c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockValueOutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_PreferredStockValueOutstanding_bb2ea530-9973-4d25-a049-d8284405661c" xlink:type="arc" order="18"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AdditionalPaidInCapital_f3709fdf-1f7c-4ada-a056-b8282c15bb35" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AdditionalPaidInCapital"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_AdditionalPaidInCapital_f3709fdf-1f7c-4ada-a056-b8282c15bb35" xlink:type="arc" order="19"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccumulatedOtherComprehensiveIncomeLossNetOfTax_e0c1898a-324d-44aa-bbff-65cd7aee4bc7" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_AccumulatedOtherComprehensiveIncomeLossNetOfTax"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_AccumulatedOtherComprehensiveIncomeLossNetOfTax_e0c1898a-324d-44aa-bbff-65cd7aee4bc7" xlink:type="arc" order="20"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_RetainedEarningsAccumulatedDeficit_2a49018c-efe3-4449-9787-e78945a11158" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_RetainedEarningsAccumulatedDeficit"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_RetainedEarningsAccumulatedDeficit_2a49018c-efe3-4449-9787-e78945a11158" xlink:type="arc" order="21"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TreasuryStockValue_b980f282-a38c-4bd2-a921-a2ca6b0b0595" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TreasuryStockValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_TreasuryStockValue_b980f282-a38c-4bd2-a921-a2ca6b0b0595" xlink:type="arc" order="22"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StockholdersEquity_c9cc20e8-416a-4a8e-a059-01d3b4b8e337" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StockholdersEquity"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_StockholdersEquity_c9cc20e8-416a-4a8e-a059-01d3b4b8e337" xlink:type="arc" order="23"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilitiesAndStockholdersEquity_40d01732-969c-4933-abb2-e0b96d6d48f0" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_LiabilitiesAndStockholdersEquity"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_us-gaap_LiabilitiesAndStockholdersEquity_40d01732-969c-4933-abb2-e0b96d6d48f0" xlink:type="arc" order="24"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ScheduleOfCondensedFinancialStatementsTable_2ac7ac40-8edb-4865-81da-e4be42b45737" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ScheduleOfCondensedFinancialStatementsTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_2131f940-9d88-482a-954d-873ddeb4eb81" xlink:to="loc_srt_ScheduleOfCondensedFinancialStatementsTable_2ac7ac40-8edb-4865-81da-e4be42b45737" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesAxis_6e848540-7c08-492a-b2c6-8c14bc26051c" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_srt_ScheduleOfCondensedFinancialStatementsTable_2ac7ac40-8edb-4865-81da-e4be42b45737" xlink:to="loc_srt_ConsolidatedEntitiesAxis_6e848540-7c08-492a-b2c6-8c14bc26051c" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesDomain_6e848540-7c08-492a-b2c6-8c14bc26051c_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_ConsolidatedEntitiesAxis_6e848540-7c08-492a-b2c6-8c14bc26051c" xlink:to="loc_srt_ConsolidatedEntitiesDomain_6e848540-7c08-492a-b2c6-8c14bc26051c_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesDomain_cc141188-24d6-4bde-9237-41164cb71768" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_ConsolidatedEntitiesAxis_6e848540-7c08-492a-b2c6-8c14bc26051c" xlink:to="loc_srt_ConsolidatedEntitiesDomain_cc141188-24d6-4bde-9237-41164cb71768" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ParentCompanyMember_310e6a84-862f-41d8-b2c0-0e8d1c62bd21" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ParentCompanyMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_ConsolidatedEntitiesDomain_cc141188-24d6-4bde-9237-41164cb71768" xlink:to="loc_srt_ParentCompanyMember_310e6a84-862f-41d8-b2c0-0e8d1c62bd21" xlink:type="arc" order="0"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/CondensedConsolidatedBalanceSheetParentheticals" xlink:type="simple" xlink:href="trup-20201231.xsd#CondensedConsolidatedBalanceSheetParentheticals"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/CondensedConsolidatedBalanceSheetParentheticals" xlink:type="extended" id="i76f3c7ae7f4948a4bdb9ab6e817ab909_CondensedConsolidatedBalanceSheetParentheticals">
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockParOrStatedValuePerShare_b797ac56-84ae-49c9-b452-da2b80869db5" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockParOrStatedValuePerShare"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_CommonStockParOrStatedValuePerShare_b797ac56-84ae-49c9-b452-da2b80869db5" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockSharesAuthorized_f263fe12-2769-4417-9374-dad39c9ffca5" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockSharesAuthorized"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_CommonStockSharesAuthorized_f263fe12-2769-4417-9374-dad39c9ffca5" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockSharesIssued_c5b9c8c0-29f6-494e-aca2-2f79ce5bea25" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockSharesIssued"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_CommonStockSharesIssued_c5b9c8c0-29f6-494e-aca2-2f79ce5bea25" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockSharesOutstanding_ab944797-bddf-41aa-9bc2-1283deac1a37" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockSharesOutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_CommonStockSharesOutstanding_ab944797-bddf-41aa-9bc2-1283deac1a37" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockParOrStatedValuePerShare_e13e325d-9934-496f-b353-393d28e86398" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockParOrStatedValuePerShare"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_PreferredStockParOrStatedValuePerShare_e13e325d-9934-496f-b353-393d28e86398" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockSharesAuthorized_43fc904c-e9b1-4b4c-b805-cd7fb8c44f44" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockSharesAuthorized"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_PreferredStockSharesAuthorized_43fc904c-e9b1-4b4c-b805-cd7fb8c44f44" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockSharesIssued_1136c76b-d5cf-4377-a1b1-0cb382a7d6b0" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockSharesIssued"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_PreferredStockSharesIssued_1136c76b-d5cf-4377-a1b1-0cb382a7d6b0" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockSharesOutstanding_6eea1089-5fb4-4510-a8a9-c5cef46d40f4" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockSharesOutstanding"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_PreferredStockSharesOutstanding_6eea1089-5fb4-4510-a8a9-c5cef46d40f4" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TreasuryStockShares_8e2d6321-90ac-4684-9fb4-1c121badf9f0" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TreasuryStockShares"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_TreasuryStockShares_8e2d6321-90ac-4684-9fb4-1c121badf9f0" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementTable_4f38333b-b26c-47ed-bd6a-62a48ea1b552" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_us-gaap_StatementLineItems_db2bbb46-96d5-40ce-82fc-b798528b68f5" xlink:to="loc_us-gaap_StatementTable_4f38333b-b26c-47ed-bd6a-62a48ea1b552" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesAxis_570df517-939c-4acd-8dab-ec60687fbb43" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_StatementTable_4f38333b-b26c-47ed-bd6a-62a48ea1b552" xlink:to="loc_srt_ConsolidatedEntitiesAxis_570df517-939c-4acd-8dab-ec60687fbb43" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesDomain_570df517-939c-4acd-8dab-ec60687fbb43_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_ConsolidatedEntitiesAxis_570df517-939c-4acd-8dab-ec60687fbb43" xlink:to="loc_srt_ConsolidatedEntitiesDomain_570df517-939c-4acd-8dab-ec60687fbb43_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesDomain_e38303c2-df67-4361-b11e-7693be57ec0a" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_ConsolidatedEntitiesAxis_570df517-939c-4acd-8dab-ec60687fbb43" xlink:to="loc_srt_ConsolidatedEntitiesDomain_e38303c2-df67-4361-b11e-7693be57ec0a" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ParentCompanyMember_b3ef3da6-e60d-4357-bed1-119a2f594cd5" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ParentCompanyMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_ConsolidatedEntitiesDomain_e38303c2-df67-4361-b11e-7693be57ec0a" xlink:to="loc_srt_ParentCompanyMember_b3ef3da6-e60d-4357-bed1-119a2f594cd5" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_StatementClassOfStockAxis_0d82063b-7be5-4e0a-beb3-8fad05c6d370" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_StatementClassOfStockAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaap_StatementTable_4f38333b-b26c-47ed-bd6a-62a48ea1b552" xlink:to="loc_us-gaap_StatementClassOfStockAxis_0d82063b-7be5-4e0a-beb3-8fad05c6d370" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_0d82063b-7be5-4e0a-beb3-8fad05c6d370_default" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_us-gaap_StatementClassOfStockAxis_0d82063b-7be5-4e0a-beb3-8fad05c6d370" xlink:to="loc_us-gaap_ClassOfStockDomain_0d82063b-7be5-4e0a-beb3-8fad05c6d370_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ClassOfStockDomain_f8a66eae-c293-4484-a5f2-4600716f4226" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ClassOfStockDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaap_StatementClassOfStockAxis_0d82063b-7be5-4e0a-beb3-8fad05c6d370" xlink:to="loc_us-gaap_ClassOfStockDomain_f8a66eae-c293-4484-a5f2-4600716f4226" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockMember_78f1d83f-fdf7-4bdb-aa24-dd35fcd64dbc" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CommonStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_f8a66eae-c293-4484-a5f2-4600716f4226" xlink:to="loc_us-gaap_CommonStockMember_78f1d83f-fdf7-4bdb-aa24-dd35fcd64dbc" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockMember_954ac26d-d813-444b-8994-8eed412707df" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PreferredStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_f8a66eae-c293-4484-a5f2-4600716f4226" xlink:to="loc_us-gaap_PreferredStockMember_954ac26d-d813-444b-8994-8eed412707df" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_TreasuryStockMember_d55d9c89-16a5-4f7a-9892-9a16085b4f8a" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_TreasuryStockMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaap_ClassOfStockDomain_f8a66eae-c293-4484-a5f2-4600716f4226" xlink:to="loc_us-gaap_TreasuryStockMember_d55d9c89-16a5-4f7a-9892-9a16085b4f8a" xlink:type="arc" order="2"/>
</link:definitionLink>
<link:roleRef roleURI="http://www.trupanion.com/role/Schedule1ParentOnlyDisclosuresScheduleCondensedStatementsofCashFlowsDetails" xlink:type="simple" xlink:href="trup-20201231.xsd#Schedule1ParentOnlyDisclosuresScheduleCondensedStatementsofCashFlowsDetails"/>
<link:definitionLink xlink:role="http://www.trupanion.com/role/Schedule1ParentOnlyDisclosuresScheduleCondensedStatementsofCashFlowsDetails" xlink:type="extended" id="ie0b93ff159fc41a68abd84160692a4d4_Schedule1ParentOnlyDisclosuresScheduleCondensedStatementsofCashFlowsDetails">
<link:loc xlink:type="locator" xlink:label="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_CondensedFinancialStatementsCaptionsLineItems"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_CashCashEquivalentsandRestrictedCashCarryingValueEndofPeriod_a6411615-9737-4ee4-8e83-cab0b71835bf" xlink:href="trup-20201231.xsd#trup_CashCashEquivalentsandRestrictedCashCarryingValueEndofPeriod"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_trup_CashCashEquivalentsandRestrictedCashCarryingValueEndofPeriod_a6411615-9737-4ee4-8e83-cab0b71835bf" xlink:type="arc" order="0"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_CashCashEquivalentsandRestrictedCashCarryingValue_12687acc-820e-4196-8c03-ff2a10717410" xlink:href="trup-20201231.xsd#trup_CashCashEquivalentsandRestrictedCashCarryingValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_trup_CashCashEquivalentsandRestrictedCashCarryingValue_12687acc-820e-4196-8c03-ff2a10717410" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ProceedsFromDividendsReceived_7901347e-bbe5-4c5d-a2a8-a51a8b93a450" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ProceedsFromDividendsReceived"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_ProceedsFromDividendsReceived_7901347e-bbe5-4c5d-a2a8-a51a8b93a450" xlink:type="arc" order="2"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetIncomeLoss_9ba86f38-7ad6-43ba-a624-45caf450cd8d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_NetIncomeLoss"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_NetIncomeLoss_9ba86f38-7ad6-43ba-a624-45caf450cd8d" xlink:type="arc" order="3"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeLossFromEquityMethodInvestments_a8d3002c-4d0f-4a5a-959a-0c91eb0d37f5" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeLossFromEquityMethodInvestments"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_IncomeLossFromEquityMethodInvestments_a8d3002c-4d0f-4a5a-959a-0c91eb0d37f5" xlink:type="arc" order="4"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncomeLossFromSubsidiariesBeforeTax_f6bbc2ac-bfd4-49bb-ad96-20341b593ceb" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncomeLossFromSubsidiariesBeforeTax"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_IncomeLossFromSubsidiariesBeforeTax_f6bbc2ac-bfd4-49bb-ad96-20341b593ceb" xlink:type="arc" order="5"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_DepreciationAmortizationAndAccretionNet_398f95d0-c831-42b5-801b-86dd25f27bde" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_DepreciationAmortizationAndAccretionNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_DepreciationAmortizationAndAccretionNet_398f95d0-c831-42b5-801b-86dd25f27bde" xlink:type="arc" order="6"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensation_8ac96506-809e-4a87-b472-66ee1af7c4fd" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ShareBasedCompensation"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_ShareBasedCompensation_8ac96506-809e-4a87-b472-66ee1af7c4fd" xlink:type="arc" order="7"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherOperatingIncomeExpenseNet_7f5f80ae-498c-48c1-bdf2-26bce31b9a2c" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_OtherOperatingIncomeExpenseNet"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_OtherOperatingIncomeExpenseNet_7f5f80ae-498c-48c1-bdf2-26bce31b9a2c" xlink:type="arc" order="8"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncreaseDecreaseInOperatingAssets_b76bf09c-1f60-463d-b13e-959095288fb9" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_IncreaseDecreaseInOperatingAssets"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_IncreaseDecreaseInOperatingAssets_b76bf09c-1f60-463d-b13e-959095288fb9" xlink:type="arc" order="9"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_f80bb13a-56aa-459c-8c3d-4d2e0e2f35e7" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_NetCashProvidedByUsedInOperatingActivities"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_f80bb13a-56aa-459c-8c3d-4d2e0e2f35e7" xlink:type="arc" order="10"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PaymentsToAcquireBusinessesNetOfCashAcquired_caab7134-0c3a-40a1-9a02-7205b0958edd" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PaymentsToAcquireBusinessesNetOfCashAcquired"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_PaymentsToAcquireBusinessesNetOfCashAcquired_caab7134-0c3a-40a1-9a02-7205b0958edd" xlink:type="arc" order="11"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ProceedsFromSaleOfEquityMethodInvestments_a95f5ef2-bcc5-4356-a434-ac6228aa3fde" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ProceedsFromSaleOfEquityMethodInvestments"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_ProceedsFromSaleOfEquityMethodInvestments_a95f5ef2-bcc5-4356-a434-ac6228aa3fde" xlink:type="arc" order="12"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PaymentsToAcquirePropertyPlantAndEquipment_e9fbc625-54d2-4e49-be90-4ec0d5a1637d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PaymentsToAcquirePropertyPlantAndEquipment"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_PaymentsToAcquirePropertyPlantAndEquipment_e9fbc625-54d2-4e49-be90-4ec0d5a1637d" xlink:type="arc" order="13"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PaymentsToAcquireInterestInSubsidiariesAndAffiliates_33c00dd3-491d-423f-a176-86816e32a9a2" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PaymentsToAcquireInterestInSubsidiariesAndAffiliates"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_PaymentsToAcquireInterestInSubsidiariesAndAffiliates_33c00dd3-491d-423f-a176-86816e32a9a2" xlink:type="arc" order="14"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ProceedsFromPaymentsForOtherFinancingActivities_77a395b2-57a0-474a-8301-c7a4a915d838" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ProceedsFromPaymentsForOtherFinancingActivities"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_ProceedsFromPaymentsForOtherFinancingActivities_77a395b2-57a0-474a-8301-c7a4a915d838" xlink:type="arc" order="15"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PaymentsForProceedsFromInvestments_bf882ef5-efc3-4123-b767-bf20ea9b1581" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PaymentsForProceedsFromInvestments"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_PaymentsForProceedsFromInvestments_bf882ef5-efc3-4123-b767-bf20ea9b1581" xlink:type="arc" order="16"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetCashProvidedByUsedInInvestingActivities_0cebabc1-bba5-4675-b9a9-4883f1d3477d" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_NetCashProvidedByUsedInInvestingActivities"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_NetCashProvidedByUsedInInvestingActivities_0cebabc1-bba5-4675-b9a9-4883f1d3477d" xlink:type="arc" order="17"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ProceedsFromIssuanceOrSaleOfEquity_574bd8f5-132a-4f63-8da4-a644e3990278" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ProceedsFromIssuanceOrSaleOfEquity"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_ProceedsFromIssuanceOrSaleOfEquity_574bd8f5-132a-4f63-8da4-a644e3990278" xlink:type="arc" order="18"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PaymentsRelatedToTaxWithholdingForShareBasedCompensation_dd145571-ff6a-44cd-8913-d78881232664" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PaymentsRelatedToTaxWithholdingForShareBasedCompensation"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_PaymentsRelatedToTaxWithholdingForShareBasedCompensation_dd145571-ff6a-44cd-8913-d78881232664" xlink:type="arc" order="19"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ProceedsFromStockOptionsExercised_7baa3188-f695-4f4e-b495-116e0349f771" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ProceedsFromStockOptionsExercised"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_ProceedsFromStockOptionsExercised_7baa3188-f695-4f4e-b495-116e0349f771" xlink:type="arc" order="20"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_ProceedsFromRepaymentsOfDebt_d563ec15-4ceb-40b7-842a-b1ccd012c9be" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_ProceedsFromRepaymentsOfDebt"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_ProceedsFromRepaymentsOfDebt_d563ec15-4ceb-40b7-842a-b1ccd012c9be" xlink:type="arc" order="21"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_RepaymentsOfLongTermDebt_a6b1abb8-942a-4ebe-b359-838b9dea8adb" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_RepaymentsOfLongTermDebt"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_RepaymentsOfLongTermDebt_a6b1abb8-942a-4ebe-b359-838b9dea8adb" xlink:type="arc" order="22"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_PaymentsOfFinancingCosts_986e6483-f59b-4005-b81d-98d302a77bbf" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_PaymentsOfFinancingCosts"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_PaymentsOfFinancingCosts_986e6483-f59b-4005-b81d-98d302a77bbf" xlink:type="arc" order="23"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetCashProvidedByUsedInFinancingActivities_7c9ab55b-94d1-4a76-8d8e-5b5abaf2568f" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_NetCashProvidedByUsedInFinancingActivities"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_NetCashProvidedByUsedInFinancingActivities_7c9ab55b-94d1-4a76-8d8e-5b5abaf2568f" xlink:type="arc" order="24"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_EffectOfExchangeRateOnCashAndCashEquivalents_2fd8de86-e745-4eac-a893-c7126a0a7639" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_EffectOfExchangeRateOnCashAndCashEquivalents"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_EffectOfExchangeRateOnCashAndCashEquivalents_2fd8de86-e745-4eac-a893-c7126a0a7639" xlink:type="arc" order="25"/>
<link:loc xlink:type="locator" xlink:label="loc_trup_CashCashEquivalentsandRestrictedCashPeriodIncreaseDecrease_b00df86a-5483-42ef-b5f6-c8875c873778" xlink:href="trup-20201231.xsd#trup_CashCashEquivalentsandRestrictedCashPeriodIncreaseDecrease"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_trup_CashCashEquivalentsandRestrictedCashPeriodIncreaseDecrease_b00df86a-5483-42ef-b5f6-c8875c873778" xlink:type="arc" order="26"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CashAndCashEquivalentsAtCarryingValue_99dd9cb4-462e-4fa0-b6e0-9599b72649ac" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CashAndCashEquivalentsAtCarryingValue"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_us-gaap_CashAndCashEquivalentsAtCarryingValue_99dd9cb4-462e-4fa0-b6e0-9599b72649ac" xlink:type="arc" order="27"/>
<link:loc xlink:type="locator" xlink:label="loc_us-gaap_CashAndCashEquivalentsAtCarryingValue_ef3fdd70-2e45-4d26-961b-3bf2b9ca9fb4" xlink:href="http://xbrl.fasb.org/us-gaap/2020/elts/us-gaap-2020-01-31.xsd#us-gaap_CashAndCashEquivalentsAtCarryingValue"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ScheduleOfCondensedFinancialStatementsTable_7485578f-c472-47d2-84eb-9d948a732d35" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ScheduleOfCondensedFinancialStatementsTable"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xlink:from="loc_srt_CondensedFinancialStatementsCaptionsLineItems_8aef62b7-4550-4d35-a514-360ce606c9d4" xlink:to="loc_srt_ScheduleOfCondensedFinancialStatementsTable_7485578f-c472-47d2-84eb-9d948a732d35" xlink:type="arc" order="1" xbrldt:closed="true" xbrldt:contextElement="segment"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesAxis_3f0aa7b0-72b5-4960-a68a-ef1a15091bf4" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesAxis"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_srt_ScheduleOfCondensedFinancialStatementsTable_7485578f-c472-47d2-84eb-9d948a732d35" xlink:to="loc_srt_ConsolidatedEntitiesAxis_3f0aa7b0-72b5-4960-a68a-ef1a15091bf4" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesDomain_3f0aa7b0-72b5-4960-a68a-ef1a15091bf4_default" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xlink:from="loc_srt_ConsolidatedEntitiesAxis_3f0aa7b0-72b5-4960-a68a-ef1a15091bf4" xlink:to="loc_srt_ConsolidatedEntitiesDomain_3f0aa7b0-72b5-4960-a68a-ef1a15091bf4_default" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ConsolidatedEntitiesDomain_4a17111a-252d-4c35-84b4-e3303dc4628e" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ConsolidatedEntitiesDomain"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srt_ConsolidatedEntitiesAxis_3f0aa7b0-72b5-4960-a68a-ef1a15091bf4" xlink:to="loc_srt_ConsolidatedEntitiesDomain_4a17111a-252d-4c35-84b4-e3303dc4628e" xlink:type="arc" order="1"/>
<link:loc xlink:type="locator" xlink:label="loc_srt_ParentCompanyMember_c9d066b4-620b-47d7-91e7-39d25b4c3415" xlink:href="http://xbrl.fasb.org/srt/2020/elts/srt-2020-01-31.xsd#srt_ParentCompanyMember"/>
<link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srt_ConsolidatedEntitiesDomain_4a17111a-252d-4c35-84b4-e3303dc4628e" xlink:to="loc_srt_ParentCompanyMember_c9d066b4-620b-47d7-91e7-39d25b4c3415" xlink:type="arc" order="0"/>
</link:definitionLink>
</link:linkbase>

9 Subsequent Filings that Reference this Filing

  As Of               Filer                 Filing    For·On·As Docs:Size             Issuer                      Filing Agent

 5/05/23  Trupanion, Inc.                   10-Q        3/31/23   72:6.6M
 2/16/23  Trupanion, Inc.                   10-K       12/31/22  104:12M
11/04/22  Trupanion, Inc.                   10-Q        9/30/22   74:7.1M
 8/04/22  Trupanion, Inc.                   10-Q        6/30/22   71:6.4M
 4/29/22  Trupanion, Inc.                   10-Q        3/31/22   70:6.7M
 2/17/22  Trupanion, Inc.                   10-K       12/31/21  104:11M
11/04/21  Trupanion, Inc.                   10-Q        9/30/21   64:5.8M
 8/06/21  Trupanion, Inc.                   10-Q        6/30/21   64:6.1M
 4/30/21  Trupanion, Inc.                   10-Q        3/31/21   64:5.2M

12 Previous Filings that this Filing References

  As Of               Filer                 Filing    For·On·As Docs:Size             Issuer                      Filing Agent

 2/04/21  Trupanion, Inc.                   8-K:5,9     1/29/21   14:487K
10/29/20  Trupanion, Inc.                   8-K:1,3,9  10/26/20   16:869K
 2/14/20  Trupanion, Inc.                   10-K       12/31/19   98:13M
 5/03/19  Trupanion, Inc.                   10-Q        3/31/19   74:5.6M
 2/14/19  Trupanion, Inc.                   10-K       12/31/18  102:12M
 2/14/18  Trupanion, Inc.                   10-K       12/31/17   89:11M
 2/15/17  Trupanion, Inc.                   10-K       12/31/16  102:14M
 6/03/16  Trupanion, Inc.                   8-K:5,9     5/31/16    2:66K
 5/06/16  Trupanion, Inc.                   10-Q        3/31/16   53:3.9M
 2/24/15  Trupanion, Inc.                   10-K       12/31/14  103:15M
 8/28/14  Trupanion, Inc.                   10-Q        6/30/14   60:6.7M
 6/16/14  Trupanion, Inc.                   S-1                   26:10M                                    Donnelley … Solutions/FA
↑Top
Filing Submission 0001371285-21-000041   –   Alternative Formats (Word / Rich Text, HTML, Plain Text, et al.)
Copyright © 2024 Fran Finnegan & Company LLC – All Rights Reserved.
About — Privacy — Redactions — Help — Thu., May 16, 6:47:10.2pm ET