Adherent Self-Renewable Human Embryonic Stem Cell-Derived Neural Stem Cell Line Functional Engraftment in Experimental Stroke Model 英文参考文献.docVIP

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Adherent Self-Renewable Human Embryonic Stem Cell-Derived Neural Stem Cell Line Functional Engraftment in Experimental Stroke Model 英文参考文献.doc

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Adherent Self-Renewable Human Embryonic Stem Cell-Derived Neural Stem Cell Line Functional Engraftment in Experimental Stroke Model 英文参考文献

AdherentSelf-RenewableHumanEmbryonicStemCell- DerivedNeuralStemCellLine:FunctionalEngraftmentin ExperimentalStrokeModel MarcelM.Daadi*,Anne-LiseMaag,GaryK.Steinberg DepartmentofNeurosurgeryandStanfordStrokeCenter,StanfordUniversitySchoolofMedicine,Stanford,California,UnitedStatesofAmerica Abstract Background:Humanembryonicstemcells(hESCs)offeravirtuallyunlimitedsourceofneuralcellsforstructuralrepairin neurologicaldisorders,suchasstroke.NeuralcellscanbederivedfromhESCseitherbydirectenrichment,orbyisolating specificgrowthfactor-responsiveandexpandablepopulationsofhumanneuralstemcells(hNSCs).Studieshaveindicated that the direct enrichment method generates a heterogeneous population of cells that may contain residual undifferentiatedstemcellsthatcouldleadtotumorformationinvivo. Methods/PrincipalFindings:WeisolatedanexpandableandhomogenouspopulationofhNSCs(namedSD56)fromhESCs using a defined media supplemented with epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and leukemiainhibitorygrowthfactor(LIF).ThesehNSCsgrewasanadherentmonolayerculture.Theywerefullyneuralizedand uniformlyexpressedmolecularfeaturesofNSCs,includingnestin,vimentinandradialglialmarkers.ThesehNSCsdidnot expressthepluripotencymarkersOct4orNanog,nordidtheyexpressmarkersforthemesodermorendodermlineages. Theself-renewalpropertyofthehNSCswascharacterizedbyapredominantsymmetricalmodeofcelldivision.TheSD56 hNSCsdifferentiatedintoneurons,astrocytesandoligodendrocytesthroughoutmultiplepassagesinvitro,aswellasafter transplantation.Together,thesecriteriaconfirmthedefinitiveNSCidentityoftheSD56cellline.Importantly,theyexhibited nochromosomeabnormalitiesanddidnotformtumorsafterimplantationintoratischemicbrainsandintona?¨venuderat brainsandflanks. Furthermore, hNSCsisolated undertheseconditionsmigratedtowardtheischemia-injuredadultbrain parenchymaandimprovedtheindependentuseofthestroke-impairedforelimbtwomonthspost-transplantation. Conclusions/Significance:TheSD56humanneuralstemcellsderive