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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 culture conditions and differentiation in vitro of human embryonic neural stem cells. METHODS: The cells were isolated from the 12-week-old human embryonic cortex and cultured in serum-free culture in combination with basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) and recombinant human leukemia inhibitory factor (rhLIF) The proliferation of cells was detected by BrdU labeling, and the differentiation of cells was detected by indirect immunofluorescence staining. Results: A large number of NSCs cultured in suspension could be subcultured. BrdU positive cells could differentiate into neurons, astrocytes and oligodendrocytes. Conclusion: Human embryonic cerebral cortex cells isolated from cell culture have the basic characteristics of neural stem cells, and can be further used in basic and clinical studies.