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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 ,bFGF和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Musashil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 在体外的培养条件下 ,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 culture conditions and passage of human neural stem cells in vitro. Methods The neural cells were isolated from the fetal brain by mechanical method, cultured in N2 medium, bFGF and EGF were used to stimulate the cell proliferation. The traditional method and the method of dividing the neurospheres were subcultured. Immunohistochemical staining of cultured cells and Their differentiated cells were identified. Results The human neural stem cells were successfully cultured from the fetal brain and grew in suspension in the culture condition to form neurospheres. Most of the cells expressed vimentin and markers of Musashil, two types of neural stem cells. These cells can differentiate into nerve There were a few oligodendrocytes cultured in early stage. In this culture condition, the growth of neural stem cells was slower, and the method of cutting the neurospheres kept the relationship between cells. Neural stem cells Can get a larger rate of amplification. Conclusion Under the culture conditions in vitro, neural stem cells can be cultured from fetal brain tissue, which can be used as a potential source of cells for the treatment of CNS diseases.