人类神经干细胞的长期培养和传代

The long-term culture and passaging of human neural stem cells

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 explore the culture conditions for human neural stem cells and to investigate the passaging method.Methods The cells from the embryonic human cortex were mechanically dissociated.N2 medium was adapted to culture the cells, bFGF and EGF were added to expand the cells, The cells were identified by immunocytochemistry.Results Neural stem cells from embryonic humans have been successfully cultured. They formed typical neurospheres in suspension,and the majorities of the cells expressed vimentin and Musashil,which were the markers for neural stem cells. The cultured cell could differentiated into neurons and astrocytes. The neural stem cells multiplied very slowly under the culture conditions, however the best expansion was achieved when the neurospheres were disected into several parts and the cell link was conserved when passaging.Conclusions Human neural stem cells could be cultured from embryonic brains.They could form the typical neurospheres in suspension in vitro, which may be potential source for transplantation in treating CNS disorders in humans.