Efficient Generation of Fully Reprogrammed Human iPS Cells via Polycistronic Retroviral Vector and a New Cocktail of Chemical Compounds 英文参考文献.docVIP

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Efficient Generation of Fully Reprogrammed Human iPS Cells via Polycistronic Retroviral Vector and a New Cocktail of Chemical Compounds 英文参考文献.doc

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Efficient Generation of Fully Reprogrammed Human iPS Cells via Polycistronic Retroviral Vector and a New Cocktail of Chemical Compounds 英文参考文献

EfficientGenerationofFullyReprogrammedHumaniPS CellsviaPolycistronicRetroviralVectorandaNew CocktailofChemicalCompounds ZhonghuiZhang1,3,YongxingGao1,AlbertGordon1,ZackZ.Wang1,ZhijianQian2,Wen-ShuWu1,3* 1StemandProgenitorCellBiologyProgram,CenterforMolecularMedicine,MaineMedicalCenterResearchInstitute,MaineMedicalCenter,Scarborough,Maine,United StatesofAmerica,2SectionofHematology/Oncology,DepartmentofMedicine,UniversityofIllinoisatChicago,Chicago,Illinois,UnitedStatesofAmerica,3Children’s HospitalOaklandResearchInstitute,Oakland,California,UnitedStatesofAmerica Abstract Directreprogrammingofhumansomaticcellsintoinducedpluripotentstem(iPS)cellsbydefinedtranscriptionfactors(TFs) providesgreatpotentialforregenerativemedicineandbiomedicalresearch.Thisprocedurehasmanychallenges,including lowreprogrammingefficiency,manypartiallyreprogrammedcolonies,somaticcodingmutationsinthegenome,etc.Here, wedescribeasimpleapproachforgeneratingfullyreprogrammedhumaniPScellsbyusingasinglepolycistronicretroviral vectorexpressingfourhumanTFsinasingleopenreadingframe(ORF),combinedwithacocktailcontainingthreesmall molecules(Sodiumbutyrate,SB431542,andPD0325901).OurresultsdemonstratethathumaniPScellsgeneratedbythis approachexpresshumanEScellsmarkersandexhibitpluripotencydemonstratedbytheirabilitiestodifferentiateintothe threegermlayersinvitroandinvivo.Notably,thisapproachnotonlyprovidesamuchfasterreprogrammingprocessbut also significantly diminishes partially reprogrammed iPS cell colonies, thus facilitating efficient isolation of desired fully reprogrammediPScellcolonies. Citation:ZhangZ,GaoY,GordonA,WangZZ,QianZ,etal.(2011)EfficientGenerationofFullyReprogrammedHumaniPSCellsviaPolycistronicRetroviralVector andaNewCocktailofChemicalCompounds.PLoSONE6(10):e26592.doi:10.1371/journal.pone.0026592 Editor:JialinCharlesZheng,UniversityofNebraskaMedicalCenter,UnitedStatesofAmerica ReceivedApril19,2011;AcceptedSeptember29,2011;PublishedOctober26,2011 Copyright: ? 2011 Zhang et al. This is an open-