人胎脑皮质神经干细胞的分离培养及鉴定

Separation and identification of neural stem cells from human fetal cerebral cortex in vitro

目的探讨人脑皮质中分离培养神经干细胞并鉴定其增殖及分化潜能。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30周自愿水囊引产人胎脑皮质中分离出神经干细胞,并进行培养、传代、分化观察,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30周人胎脑皮质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,表达神经巢蛋白抗原(Nestin);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和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;30周人胎脑皮质仍能培养出具有自我复制和分化潜能的神经干细胞。

Objective To separate neural stem cells from human fetal cerebral cortex of 30 weeks and their proliferation capacity as well as differentiating potential.Methods Human fetal cerebral cortex at embryonic age 30 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 tecnique,then identified for The cells from a clone were cultured passed and differentiated.Cultured and differentiated cells were identified with immunocytochemistry staining.Results The human neural stem cells having the abilities of self renewal and multipotency were success fully isolated and cultured from human fetal cerebral cortex with serum free midium.They formed typical neuraospheres in suspension.The cells could be cloned and passed continuously,expressing Nestin antigen.serum midium 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 30 from human fetal cerebral cortex remain to be cultured in vitro.Which have the abilities of self renewal and multipotencypotential.