人脑神经干细胞体外培养中增殖、自我更新及多向分化潜能的观察

背景神经干细胞为神经系统损伤的修复开辟了新的途径,也是发现某些细胞因子和其作用机制的良好模型,神经干细胞的体外培养、增殖和诱导分化的探讨是这些研究的重要基础. 目的分离培养和鉴定人脑神经干细胞. 设计和方法采用无血清培养基分离和培养人胚胎脑皮质分离和培养神经干细胞,并应用免疫荧光技术进行细胞自我更新能力、巢蛋白表达和多向分化潜能的鉴定. 地点和材料实验在福建医科大学分子医学研究中心实施.实验材料为自然流产的孕龄 6～ 8周的人胚胎脑组织. 主要观察指标人脑神经干细胞在体外培养条件中的增殖、自我更新能力、巢蛋白表达和多向分化潜能. 结果人胚胎脑组织分离培养的细胞具有神经干细胞的特征,并可在体外大量增殖,经诱导可分化为神经元、星形胶质细胞和少突胶质细胞. 结论从人胚胎脑皮质成功分离培养出的神经干细胞,是研究神经干细胞诱导分化的良好模型.

Observation on the proliferation,self-renewal ability and multipotency of differentiation of nerve stem cells isolated from human brain during culture in vitro

BACKGROUND:Neural stem cells(NSCs) have raised hopes that researchers can find ways to repair nervous system damage.NSCs cultured in vitro can be a good model to discover some cytokines and explore their mechanisms.The studies on culture,proliferation,and inducing differentiation of neural stem cells in vitro are basis for these works. OBJECTIVE:To isolate,culture and characterize the neural stem cells from human brain. DESIGN and METHODS:Neural stem cells from human embryonic cerebral cortex were isolated and cultured by using serum-free medium.Their self-renewal ability,expression of nestin and multipotency of differentiation were identified with immunofluorescence staining. SETTING and PARTICIPANTS:The experiment was performed in Research Center of Molecular Medicine in Fujian Medical University. Human fetal(spontaneous abortion in gestation weeks 6-8) brain tissues were obtained from the Departments of Obstetrics in the First Affiliated Hospital of Fujian Medical University and Fuzhou General Hospital of Nanjing Military Region. MAIN OUTCOME MEASURES:The capability of proliferation and survival of neural stem cells were studied in vitro.The self-renewal ability,expression of nestin and multipotency of differentiation of neural stem cells were studied,too. RESULTS:NSCs were isolated from human embryonic cerebral cortex by serum-free medium and capable to proliferation and long-term survival in vitro.The cells could be differentiated into neurons,astrocytes,and oligodendrocytes. CONCLUSION:NSCs are successfully isolated and cultured from human embryonic cerebral cortex.They are good models for research of inducing differentiation.