人胚神经干细胞的体外培养及鉴定

Culture and identification of human embryonic neural stem cells in vitro

目的探讨人胚神经干细胞的体外培养和诱导分化的条件。方法从药物流产的12周到16周的人胚胎海马组织中分离神经干细胞,在EGF、bFGF和LIF联合作用下使其稳定增殖,并用10%的胎牛血清诱导其贴壁分化,应用免疫荧光染色方法行Nestin、NSE、MAP-2、GFAP和GalC免疫荧光染色,对神经干细胞及其分化的细胞进行鉴定。结果体外培养的神经干细胞增殖成神经干细胞球并传代,鉴定为Nestin染色阳性细胞,并可诱导分化为神经细胞、星形胶质细胞和少突胶质细胞。结论利用无血清培养技术和特定生长因子,可培养出在体外稳定增殖并有多向分化潜能的人胚神经干细胞。

Objective To explore the condition of cultivating human embryonic neural stem cells in vitro and its identification. Methods Obtain human neural stem cells from hippocampal tissue of drug aborted embryo aged from 12 to 16 weeks and proliferate it steadily with EGF.bFGF and LIF. Meanwhile, to induce human neural stem cell sphere differentiation and identify it with measure of immunofluorescence including Nestin, NSE, MAP-2, GFAP and GalC. Results Cultivated neural stem cells proliferated into sphere and was Nestin positive. The cells in the sphere could express specific antigens of neurons, astrocytes and oligodendrocyte after differentiation. Conclusion With the help of growth factors and no-serum technique, human embryonic neural stem cells which have multiple potency of differentiation could proliferate in vitro steadily.