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【目的】探讨人类神经干细胞的体外培养条件及其传代的方法。【方法】采用机械方法从胎脑中分离神经细胞 ,应用N2培养基进行培养 ,碱性成纤维细胞生长因子 (bFGF)和表皮生长因子 (EGF)刺激细胞扩增 ;传统方法和对神经球切割的方法进行传代培养 ;应用免疫组织化学染色对培养的细胞及其分化的细胞进行鉴定。【结果】从流产胎脑当中成功培养出人类的神经干细胞 ,培养条件下呈悬浮状态生长 ,形成神经球 ,绝大多数的细胞表达波形蛋白和Musashi1两种神经干细胞的标志物 ;这种细胞可分化为神经元和星型胶质细胞 ,早期的培养有少量的少突胶质细胞 ;在这种培养条件下 ,神经干细胞生长速度较慢 ,而采用切割神经球的方法保持了细胞间的联系 ,神经干细胞可获得较大的扩增速度。【结论】体外的培养条件下 ,可从胎脑组织中培养出神经干细胞 ,它可做为中枢神经系统疾病移植治疗的潜在细胞来源。
【Objective】 To investigate the in vitro culture conditions of human neural stem cells and their passage methods. 【Methods】 Nerve cells were isolated from fetal brain by mechanical method, and cultured in N2 medium. Basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) were used to stimulate cell proliferation. Conventional methods and excision of neurospheres Methods: The cultured cells and their differentiated cells were identified by immunohistochemical staining. 【Results】 Human neural stem cells were successfully cultured from aborted fetal brain and grew in suspension under culture conditions to form neurospheres. Most of the cells expressed vimentin and Musashi1 markers of neural stem cells; Differentiation into neurons and astrocytes, the early culture of a small amount of oligodendrocytes; in this culture conditions, neural stem cells grow slower, and the use of cutting the ball to maintain the way the relationship between the cells , Neural stem cells can obtain a larger rate of amplification. 【Conclusion】 Neural stem cells can be cultured from fetal brain tissue under in vitro culture conditions, which can be used as a potential source of cells for the treatment of CNS diseases.