人胚胎神经干细胞的分离培养及鉴定研究

Isolation,cultivation and characterization of neural stem cell from human fetal cerebral cortex

目的:研究人胚胎神经干细胞的培养条件及体外分化情况。方法:从12周龄人胚胎脑皮质分离细胞,采用无血清培养技术,协同应用碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)和重组人白血病抑制因子(rhLIF)进行培养;5-溴脱氧尿嘧啶核苷(BrdU)标记检测细胞的增殖能力,间接免疫荧光化学法检测细胞的分化情况。结果:培养得到的大量半悬浮生长的神经干细胞球能够传代培养;BrdU标记阳性,可诱导分化为神经元、星形胶质细胞和少突胶质细胞。结论:人胚胎脑皮质分离细胞培养得到的细胞群具有神经干细胞的基本特征,可进一步用于基础及临床研究。

Objective:To study the culturing condition and in vitro differentiation of human neural stem cell.Method:The cells isolated from fresh human fetal cerebral cortex(12weeks post conception)were seeded into defined serum-free medium containing basic fibroblast growth factor(bFGF),epidermal growth factor(EGF)and recombinant human leukemia inhibitory factor(rhLIF).The proliferative ability of these cells were tested by BrdU labeling.The cells were induced to differentiation by exposure to fetal calf serum and identified with indirect immunoflurenscence cytochemistry.Result:The harvested cells growed as neurosphere in suspension and passed serially.The cells in the sphere could incorporate BrdU and expressed specific antigens of neurons,astrocytes and oligodendrocytes after differentiation.Conclusion:Our cell line has the common features of neural stem cell and can be used for further basic research and clinical application.