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目的:建立人胚神经干细胞(humanNeuralStemCells,hNSC)的体外培养体系。方法:选取3个月～4个月大的引产胎儿,分离原代人胚神经干细胞,添加碱性成纤维细胞生长因子(bFGF)、N2促进hNSC的增殖,使用免疫细胞化学方法与诱导分化实验鉴定其神经干细胞的特性。结果:成功培养出人胚神经干细胞,原代培养的hNSC悬浮生长,似桑椹状,表达巢蛋白(Nestin)抗原,诱导分化后可表达神经元与神经胶质细胞特异抗原βⅢ与神经胶质细胞特异抗原GFAP,并能自我增殖,目前已成功传代10余次(5月余)。结论:使用本方法可以获得大量纯化hNSC,为临床实验提供适合的移植材料。
Objective: To establish an in vitro culture system of human neural stem cells (hNSCs). METHODS: Primary human embryonic neural stem cells were isolated from 3 months to 4 months old. Basic fibroblast growth factor (bFGF) and N2 were added to promote hNSC proliferation. Immunocytochemistry and differentiation assay Identification of the characteristics of neural stem cells. Results: Human embryonic neural stem cells were successfully cultured. Primary cultured hNSCs grew in suspension and appeared mulberry like and expressed Nestin antigen. After inducing differentiation, hNSC could express neurons and glial cell specific antigen β Ⅲ and glial cells Specific antigen GFAP, and can self-proliferation, has been successfully passaged more than 10 times (May more than). Conclusion: The method can be used to obtain a large number of purified hNSC, for clinical trials to provide suitable materials for transplantation.