Direct Reprogramming of Rat Neural Precursor Cells and Fibroblasts into Pluripotent Stem Cells 英文参考文献.docVIP

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Direct Reprogramming of Rat Neural Precursor Cells and Fibroblasts into Pluripotent Stem Cells 英文参考文献.doc

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Direct Reprogramming of Rat Neural Precursor Cells and Fibroblasts into Pluripotent Stem Cells 英文参考文献

DirectReprogrammingofRatNeuralPrecursorCellsand FibroblastsintoPluripotentStemCells Mi-YoonChang1,DohoonKim1,Chun-HyungKim1,Hoon-ChulKang1¤,EungiYang1,Jung-IlMoon1, SanghyeokKo1,JunpilPark1,Kyung-SoonPark3,Kyung-AhLee3,Dong-YounHwang3,YoungChung4, RobertLanza4,Kwang-SooKim1,2,3* 1Molecular Neurobiology Laboratory, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, United States of America, 2Harvard Stem Cell Institute, Cambridge,Massachusetts,UnitedStatesofAmerica,3CHAStemCellInstitute,PochonCHAUniversityCollegeofMedicine,Seoul,Korea,4StemCellInternational,Inc., Worcester,Massachusetts,UnitedStatesofAmerica Abstract Background: Given the usefulness of rats as an experimental system, an efficient method for generating rat induced pluripotent stem(iPS) cells would provide researchers with apowerful tool for studyinghuman physiology and disease. Here,wereportdirectreprogrammingofratneuralprecursor(NP)cellsandratembryonicfibroblasts(REF)intoiPScellsby retroviraltransductionusingeitherthree(Oct3/4,Sox2,andKlf4),four(Oct3/4,Sox2,Klf4,andc-Myc),orfive(Oct3/4,Sox2, Klf4,c-Myc,andNanog)genes. MethodologyandPrincipalFindings:iPScellsweregeneratedfrombothNPandREFusingonlythree(Oct3/4,Sox2,and Klf4)geneswithoutc-Myc.TwofactorswerefoundtobecriticalforefficientderivationandmaintenanceofratiPScells:theuse ofratinsteadofmousefeeders,andtheuseofsmallmoleculesspecificallyinhibitingmitogen-activatedproteinkinaseand glycogen synthase kinase 3 pathways. In contrast, introduction of embryonic stem cell (ESC) extracts induced partial reprogramming,butfailedtogenerateiPScells.However,whencombinedwithretroviraltransduction,thismethodgenerated iPScellswithsignificantlyhigherefficiency.Morphology,geneexpression,andepigeneticstatusconfirmedthattheseratiPS cellsexhibitedESC-likeproperties,includingtheabilitytodifferentiateintoallthreegermlayersbothinvitroandinteratomas. Inparticular,wefoundthattheseratiPScellscoulddifferentiatetomidbrain-likedopamineneuronswithahighefficiency. C