人类神经干细胞的长期培养和传代

【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。

The Long-Term Culture and Passaging of Human Neural Stem Cells

Objectives] To explore the culture conditions for human neural stem cells and to investigate the passaging method The cells from the embryonic human brain cortiex were mechanically dissociated Cells were cultured in N2 medium with bFGF and EGF ,and identified by immunocytochemistry Neural stem cells from abortive embryonic humans had been successfully cultured Typical neurospheres were developed in suspension, and majorities of the cells expressed vimentin and Musashil, which were the markers for neural stem cells The cells could differentiate into neurons and astrocytes Neural stem cells multiplied very slowly under the culture conditions, and the best expansion was achieved when the neurospheres were disected into several parts and the cell link was conserved when passaging [Conclusion] Human neural stem cells can be cultured from embryonic brains They formed the typical neurospheres in suspension in vitro, which serves as potential source for transplantation in treating CNS disorders in humans