人胎脑神经干细胞的体外分离、培养与鉴定

发布时间：2018-11-29 14:07

【摘要】：目的探索人胎脑神经干细胞的体外分离培养条件,从而在体外大量增殖神经干细胞,并观察神经干细胞增殖和分化的特点。方法采用含碱性成纤维细胞生长因子(bFGF)和表皮细胞生长因子(EGF)的无血清培养技术,从14周自愿水囊引产人胎脑海马皮质中分离出神经干细胞,并进行培养、传代、分化观察,应用免疫荧光细胞化学技术对培养的细胞及其分化的细胞进行鉴定。结果从14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,该细胞具有连续增殖能力,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。结论在含有EGF和bFGF的无血清培养液中,从人胎脑能分离培养出具有自我复制和分化潜能的神经干细胞,并能在体外扩增,可进一步用于基础及临床研究。
[Abstract]:Objective to explore the conditions of isolation and culture of neural stem cells from human fetal brain in vitro, so as to proliferate neural stem cells in vitro and observe the characteristics of proliferation and differentiation of neural stem cells. Methods using serum-free culture technique containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF), neural stem cells (NSCs) were isolated from the cortex of human fetal brain induced by 14 weeks of voluntary water sac, and were cultured and subcultured. The cultured cells and the differentiated cells were identified by immunofluorescence cytochemistry. Results the neural stem cells with self-renewal and multi-differentiation potential were successfully isolated from the human fetal cerebral cortex at the 14th week. The cells grew in suspension state and formed a neurosphere in serum-free culture. The cells had the ability of continuous proliferation and could be subcultured. Expression of nestin antigen (Nestin); The differentiation of neural stem cells was induced by serum-containing culture. The differentiated cells expressed specific antigens of neuronal cells and glial cells. Conclusion in serum-free medium containing EGF and bFGF, neural stem cells with self-replication and differentiation potential can be isolated from human fetal brain and can be amplified in vitro, which can be further used in basic and clinical research.
【作者单位】：安徽医科大学附属安庆医院神经外科;安庆师范学院生命科学系;
【分类号】：R329

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