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《Journal of Harbin Medical University》 2007-04
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Differentiation of Kunming mouse embryonic stem cells into neuronal stem cells in vitro

BAO Jun-hong,LIU Hui-wen,WANG Jin-xiu (Department of Histology and Embryology,Harbin Medical University,Harbin 150081,China)  
Objective To develop an efficient method by which embryonic stem cells(ESC) can be cultured and induced to differentiate towards neuronal stem cells(NSC) in vitro.Methods Isolated the blastocysts from 3.5d pregnant Kunming mouse.The inner cell mass(ICM) which were obtained following hatch were cultured on the mouse embryonic fibroblast cell(MEF) feeder layer.The stem cells were identified by alkaline phosphatase staining,differentiation ability in vitro and Oct-4 immunocytochemical staining.ESC was subjected to an 8d induction procedure which consisted of 4d of culture as embryonic bodies(EB) without all-trans retinoic acid(RA) and hLIF followed by 4d of culture in the presence of RA.Nestin immunocytochemical staining was performed after amplification in NSC screening medium.Results The isolated cells were positive for Oct-4 and alkaline phosphatase.Therefore the isolated cells presented the general characters of ESC.The ESC which induced by RA was cultivated in NSC screening medium,formed a number of spherical structures.Spherical structures were positive for nestin immunocytochemical staining.Conclusion We obtain large NSC then the ESC which induced by RA were cultivated in NSC screening medium,this result provides a new pathway for healing of nervous system injury and neurodegenerative disease.
【Fund】: 黑龙江省教育厅资助项目(10551139)
【CateGory Index】: R329
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