Micro-Arrayed Human Embryonic Stem Cells-Derived Cardiomyocytes for In Vitro Functional Assay 英文参考文献.docVIP

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Micro-Arrayed Human Embryonic Stem Cells-Derived Cardiomyocytes for In Vitro Functional Assay 英文参考文献.doc

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Micro-Arrayed Human Embryonic Stem Cells-Derived Cardiomyocytes for In Vitro Functional Assay 英文参考文献

Micro-ArrayedHumanEmbryonicStemCells-Derived CardiomyocytesforInVitroFunctionalAssay ElenaSerena1,2,ElisaCimetta1,2,SusiZatti1,2,TaniaZaglia2,MonicaZagallo1,2,GordonKeller3, NicolaElvassore1,2 * 1Industrial Engineering Department, University of Padova, Padova, Italy, 2Venetian Institute of Molecular Medicine, Padova, Italy, 3McEwen Centre for Regenerative Medicine,UniversityHealthNetwork,Toronto,Ontario,Canada Abstract Introduction:Theheartisoneoftheleastregenerativeorgansinthebodyandanymajorinsultcanresultinasignificant lossofheartcells.Thedevelopmentofaninvitro-basedcardiactissuecouldbeofparamountimportanceformanyaspects ofthecardiologyresearch.Inthiscontext,wedevelopedaninvitroassaybasedonhumancardiomyocytes(hCMs)andad hocmicro-technologies,suitableforseveralapplications:frompharmacologicalanalysistophysio-phatologicalstudieson transplantablehCMs.WefocusedonthedevelopmentofanassayabletoanalyzenotonlyhCMsviability,butalsotheir functionality. Methods: hCMs were cultured onto a poly-acrylamide hydrogel with tunable tissue-like mechanical properties and organizedthroughmicropatterningina20620array.ArrayedhCMswerecharacterizedbyimmunofluorescence,GAP-FRAP analysesandliveanddeadassay.Theirfunctionalitywasevaluatedmonitoringtheexcitation-contractioncoupling. Results:MicropatternedhCMsmaintainedtheexpressionofthemajorcardiacmarkers(cTnT,cTnI,Cx43,Nkx2.5,a-actinin) and functional properties. The spontaneous contraction frequency was (0.8360.2) Hz, while exogenous electrical stimulationleadtoanincreaseupto2Hz.Asproofofconceptthatourdevicecanbeusedforscreeningtheeffectsof pathologicalconditions,hCMswereexposedtoincreasinglevelsofH2O2.Remarkably,hCMsviabilitywasnotcompromised withexposureto0.1mMH2O2,buthCMscontractilitywasdramaticallysuppressed.Asproofofconcept,wealsodeveloped amicrofluidicplatformtoselectivelytreatareasofthecellarray,intheperspectiveofperformingmulti-parametricassay. Conclusions: Such system could be a useful tool for testing the effects of multiple cond