Differentiation of neuron-like cells from mouse parthenogenetic embryonic stem cells

(整期优先)网络出版时间:2013-04-14
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Parthenogeneticembryonicstemcellshavepluripotentdifferentiationpotentials,akintofertilizedembryo-derivedembryonicstemcells.Theaimofthisstudywastocomparetheneuronaldifferentiationpotentialofparthenogeneticandfertilizedembryo-derivedembryonicstemcells.Beforedifferentiation,karyotypeanalysiswasperformed,withnormalkaryotypesdetectedinbothparthenogeneticandfertilizedembryo-derivedembryonicstemcells.SexchromosomeswereidentifiedasXX.Immunocytochemistryandquantitativereal-timePCRdetectedhighexpressionofthepluripotentgene,Oct4,atboththemRNAandproteinlevels,indicatingpluripotentdifferentiationpotentialofthetwoembryonicstemcellsubtypes.Embryonicstemcellswereinducedwithretinoicacidtoformembryoidbodies,andthendispersedintosinglecells.SinglecellsweredifferentiatedinN2differentiationmediumfor9days.Immunocytochemistryshowedparthenogeneticandfertilizedembryo-derivedembryonicstemcellsbothexpresstheneuronalcellmarkersnestin,βIII-tubulinandmyelinbasicprotein.Quantitativereal-timePCRfoundexpressionofneurogenesisrelatedgenes(Sox-1,Nestin,GABA,Pax6,Zic5andPitx1)inbothtypesofembryonicstemcells,andOct4expressionwassignificantlydecreased.NestinandPax6expressioninparthenogeneticembryonicstemcellswassignificantlyhigherthanthatinfertilizedembryo-derivedembryonicstemcells.Thus,ourexperimentalfindingsindicatethatparthenogeneticembryonicstemcellshavestrongerneuronaldifferentiationpotentialthanfertilizedembryo-derivedembryonicstemcells.