人胎脑神经干细胞的体外培养

目的从人胎脑纹状体中分离培养并鉴定神经干细胞。方法采用包含碱性成纤维细胞生长因子(bFGF)和表皮细胞生长因子(EGF)的无血清培养和单细胞克隆技术,从36周自愿水囊引产人胎脑纹状体中分离出神经干细胞,并观察神经干细胞体外培养、传代、分化潜能。结果从36周人胎脑纹状体中成功分离出具有自我更新和多分化潜能的神经干细胞,在无血清培养时细胞呈悬浮状态生长,形成神经球,该细胞具有连续克隆能力,可传代培养,呈Nestin免疫反应阳性细胞;在含血清培养时诱导神经干细胞分化,分化后的细胞表达神经元细胞、星形胶质细胞和少突胶质细胞的特异性抗原。结论神经干细胞的存活和分裂有赖于EGF和bFGF的共同作用;36周人胎脑纹状体仍能培养出具有自我复制和分化潜能的神经干细胞。

Culture of Neural Stem Cells Isolated from Human Fetal Brain in Vitro

Objective To separate,culture and identify neural stem cells from human fetal striatum.Methods Human fetal striatum at embryonic age 36 with induction of labor with water bag were collected and cultured with serum free medium containing basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF)and single cell clone technique,then cells from a clone were cultured,passed and differentiated. Cultured and differentiated cells were identified with immunocytochemistry staining.Results Human neural stem cells having the abilities of self-renewal and multipotency were successfully isolated and cultured from human fetal striatum with serum free medium.They formed typical neuraospheres in suspension.The cells could be cloned and passed continuously,expressing Nestin antigen.Serum medium induced neural stem cells to differentiate and express specific antigens of neuron,astrocyte and oligodendrocyte.Conclusion The results suggest that stock and cleavage of neural stem cells depend on cooperations of EGF and bFGF.Neural stem cells at embryonic age 36 from human fetal cerebral cortex remain to be cultured in vitro,which have the abilities of self-renewal and multipotencypotential.