人胚神经干细胞的长期培养和传代

目的 探讨人胚神经干细胞的体外培养条件及传代方法.方法 采用机械方法从胎脑中分离神经细胞,用Eagles MEM培养基,加入碱性成纤维生长因子和表皮生长因子刺激细胞增殖,传统的方法进行传代培养;应用免疫组织化学和免疫荧光染色对培养的细胞及分化的细胞进行鉴定.结果 培养第8天,细胞具备神经元形态;折光性强的锥形或圆形胞体及长的多极突起.免疫组化和免疫荧光染色,神经元特异性烯醇化酶、神经蛋白、胶质纤维酸性蛋白酶、微管相关蛋白为阳性.结论 在体外的培养条件下,可以从胎脑组织中培养出神经干细胞,为中枢神经系统疾病移植打下了基础.

The Long-term Culture and Passaging of Human-embryo Nerve Trunk cells

Objective To explore the culture conditions for human-embryo nerve trunk cells and to investigate the passaging methods.Methods To separate the nerve cell from the embryonic human embryo cortex with the mechanically methods. Eagles MEM medinum was adapted to culture the cells bFGF and EGF were added to expand the cells.The cells were cultured and passed by traditional methods. The tecknigue of immunocytochemistry and immunocy tofluorescence were applied to identify the cells.Results On the eighth day of co-culture,the cells had already the typical morphological neuronal traits:the refractile and cone-like or spherical cell bodies and long processes.Immunocytochemistry and immunocytofluorescence expressed neurom-specific enolase(NSE),NF-MGFAP and MAP2ab a marker of mature neurons.Conclusions Human nerve trunk cells could be cultured from embryonic brains.