Isolation, Characterization, and Differentiation Potential

CellTransplantation,Vol.19,pp.279–289,2010

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Copyright 2010CognizantComm.Corp.0963-6897/10$90.00+.00DOI:10.3727/096368909X481764E-ISSN1555-3892

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Isolation,Characterization,andDifferentiationPotential

ofCanineAdipose-DerivedStemCells

N.M.Vieira,*V.Brandalise,*E.Zucconi,*M.Secco,*B.E.Strauss, andM.Zatz*

*HumanGenomeResearchCenter,BiosciencesInstitute,UniversityofSa oPaulo,Sa oPaulo,Brazil

ViralVectorGroup,HeartInstitute,InCor,UniversityofSa oPaulo,Sa oPaulo,Brazil

Adiposetissuemayrepresentapotentialsourceofadultstemcellsfortissueengineeringapplicationsin

veterinarymedicine.Itcanbeobtainedinlargequantities,underlocalanesthesia,andwithminimaldiscom-

fort.Inthisstudy,canineadiposetissuewasobtainedbybiopsyfromsubcutaneousadiposetissueorby

suction-assistedlipectomy(i.e.,liposuction).Adiposetissuewasprocessedtoobtainafibroblast-likepopula-

tionofcellssimilartohumanadipose-derivedstemcells(hASCs).Thesecanineadipose-derivedstemcells

(cASCs)canbemaintainedinvitroforextendedperiodswithstablepopulationdoublingandlowlevelsof

senescence.ImmunofluorescenceandflowcytometryshowthatthemajorityofcASCsareofmesodermal

ormesenchymalorigin.cASCsareabletodifferentiateinvitrointoadipogenic,chondrogenic,myogenic,

andosteogeniccellsinthepresenceoflineage-specificinductionfactors.Inconclusion,likehumanlipoaspir-

ate,canineadiposetissuemayalsocontainmultipotentcellsandrepresentanimportantstemcellsource

bothforveterinarycelltherapyaswellaspreclinicalstudies.

Keywords:Canineadipose-derivedstemcells;Tissueengineering;Veterinarycelltherapy

INTRODUCTION

Apromisingapplicationintheemergingfieldofvet-

erinaryregenerativemedicineandsurgeryiscellther-

apy,renderingtheisolationandcharacterizationofstem

cellsfromavarietyofsourcesareasofgreatinterest.

Anabundantandaccessiblesourceofstemcellsis

adiposetissue.Thesecells,calledadipose-derivedstro-

malcells(ASCs),arefibroblast-likecellscapableof

multipotentialdifferentiation,whichhavebeenfoundin

differentspecies(4,27,29,35).Severalgroupshave

demonstratedthathumanmesenchymalcellswithinthe

stromal-vascularfraction(SVF)ofsubcutaneousadipose

tissue[processedlipoaspirate(PLA)cells]arecapable

ofdifferentiationinmultiplelineages,includingmyo-

cytes,inthepresenceoflineage-specificinductiveme-

dia(2,5,8–10,15,16,19,20,22,23,25,34,35).

Inhumans,ASCsforautologoustransplantationare

isolatedrelativelyquicklyfromadiposetissuebycolla-

genasedigestion(6).WehaverecentlyshownthatASCs

fromhumansubcutaneousfatwereabletodifferentiate

inadipogenic,osteogenic,chondrogenic,andmyogenic

lineagesandproducehumanmuscleproteinsinvitro

andinvivo(30,31).Successfultransplantationofcanineadipose-derivedstemcells(cACSs)indogswasreportedbyLietal.(12)andBlacketal.(1).However,thesemanuscriptslackedthefullcharacterizationoftheadministeredcellpopulation.Herewereporttheisolation,characteriza-tion,andmultilineagedifferentiationpotentialofcASCsfromsubcutaneousadiposetissuebyliposuctionandbi-opsyprocedures.MATERIALSANDMETHODSAllexperimentalprotocolswereapprovedbytheeth-icscommitteeonanimalusefromtheInstituteofBiosci-ences,UniversityofSa oPaulo.Forthisstudy,adiposetissuewascollectedfromnormalgoldenretrieverdogsfromtheBrazilianColonyofGoldenRetrieverMuscularDystrophy,FacultyofVeterinaryMedicineandZoo-tecny,UniversityofSa oPaulo.Subcutaneousadiposetissuewascollectedfromtheareaoverthedorsalglutealmusclesof10dogs(aged4monthsto4years).AdiposeTissueHarvestingDogsweresedateduponintramuscular(IM)injectionwithmeperidine(2mg/kg)andacetylpromazine(0.05mg/kg).TheareaoverthedorsalglutealmuscleswasReceivedMarch27,2009;finalacceptanceDecember2,2009.Onlineprepubdate:December8,2009.

AddresscorrespondencetoDr.MayanaZatz,Ph.D.,HumanGenomeResearchCenter,InstituteofBiosciences,UniversityofSa oPaulo,Ruado

´ria,SaMata o,n.106CidadeUniversita oPaulo-SP,Brasil-CEP:05508-090.Tel/Fax:(55)(11)3091-7966;E-mail:mayazatz@usp.br

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