Self-Contained Induction of Neurons from Human Embryonic Stem Cells 英文参考文献.docVIP

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Self-Contained Induction of Neurons from Human Embryonic Stem Cells 英文参考文献.doc

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Self-Contained Induction of Neurons from Human Embryonic Stem Cells 英文参考文献

Self-ContainedInductionofNeuronsfromHuman EmbryonicStemCells TsuyoshiOkuno1,2,TakashiNakayama3,NaeKonishi1,HideoMichibata1,KojiWakimoto1 ,Yutaka Suzuki1,ShinjiNito1,ToshioInaba2,ImaharuNakano4,Shin-ichiMuramatsu4,MakotoTakano5 ,Yasushi Kondo1*,NobuoInoue6 1AdvancedMedicalResearchLaboratory,MitsubishiTanabePharmaCorporation,Osaka,Japan,2DepartmentofAdvancedPathobiology,GraduateSchoolofLifeand Environmental Sciences, Osaka Prefecture University, Osaka, Japan, 3Department of Biochemistry, Yokohama City University School of Medicine, Yokohama, Japan, 4Division of Neurology, Department of Medicine, Jichi Medical University, Tochigi, Japan, 5Department of Physiology, Jichi Medical University, Tochigi, Japan, 6LaboratoryofRegenerativeNeurosciences,GraduateSchoolofHumanHealthSciences,TokyoMetropolitanUniversity,Tokyo,Japan Abstract Background: Neurons and glial cells can be efficiently induced from mouse embryonic stem (ES) cells in a conditioned medium collected from rat primary-cultured astrocytes (P-ACM). However, the use of rodent primary cells for clinical applicationsmaybehamperedbylimitedsupplyandriskofcontaminationwithxeno-proteins. Methodology/PrincipalFindings:Wehavedevelopedanalternativemethodforunimpededproductionofhumanneurons underxeno-freeconditions.Initially,neuralstemcellsinsphere-likeclusterswereinducedfromhumanES(hES)cellsafter beingculturedinP-ACMunderfree-floatingconditions.Theresultantneuralstemcellscouldcircumferentiallyproliferate undersubsequentadhesiveculture,andselectivelydifferentiateintoneuronsorastrocytesbychangingthemediumtoP- ACMorG5,respectively.ThesehEScell-derivedneuronsandastrocytescouldprocurefunctionssimilartothoseofprimary cells. Interestingly, a conditioned medium obtained from the hES cell-derived astrocytes (ES-ACM) could successfully be usedtosubstituteP-ACMforinductionofneurons.Neuronsmadebythismethodcouldsurviveinmicebrainafterxeno- transplantation. Conclusion/Significance:ByinducingastrocytesfromhEScellsinachemicallydefinedmed