EGF培养条件下NGF与BDNF对大鼠海马神经干细胞向神经元分化的差异  

Difference between NGF and BDNF on the differentiation into neurons of rat hippocampus neural stem cells in the media containing EGF

作　　者：王振宇[1] 佟雷[1] 季丽莉[1] 唐源远[1] 赵久红[1] 

机构地区：[1]中国医科大学基础医学院人体解剖学教研室,辽宁沈阳110001

出　　处：《解剖学研究》2008年第2期108-111,共4页Anatomy Research

基　　金：国家自然科学基金(30470963)

摘　　要：目的探讨在表皮生长因子(EGF)培养条件下,相同浓度神经生长因子(NGF)与脑源性神经生长因子(BDNF)对成年大鼠海马神经干细胞向神经元分化比例的差异。方法用含碱性成纤维生长因子(bFGF)、EGF、B27的无血清细胞培养技术体外培养成年大鼠海马神经干细胞,单细胞克隆后行Nestin免疫细胞化学染色,诱导分化1周,行GFAP和NSE免疫细胞化学染色;根据培养液中所加营养因子的不同将单细胞克隆传代细胞分为5组培养:EGF组、NGF组、BDNF组、EGF+NGF组、EGF+BDNF组,此5组细胞培养1周,进行NSE免疫细胞化学染色,计数阳性细胞比例后进行统计学分析。结果:单细胞克隆培养后克隆球细胞表达Nestin,诱导分化1周,细胞表达NSE、GFAP;与EGF组、NGF组、BDNF组相比,EGF+NGF组和EGF+BDNF组细胞分化为神经元的比例较高(P<0.05),其中EGF+BDNF组细胞的比例最高。结论在EGF培养条件下,BDNF促进成年大鼠海马神经干细胞向神经元分化的能力高于NGF。Objective To explore the difference of nerve growth factor （NGF） and brain-derived neurotrophic factor （BDNF） on differentiation of neural stem cells （NSCs） from hippocampus of adult rats in the media containing epidermal growth factor （EGF）. Methods The serum-free medium containing bFGF, EGF and B27 was used to culture NSCs isolated from hippocampus of rats. Immunocytochemistry for Nestin was used to identify monoclonal cells, for glial fibrillary acidic protein （GFAP） and neurone specific enolase （NSE） to identify astrocytes and neurons respectively one week after NSCs were differentiated. The passaged monoclonal NSCs were divided into five groups： EGF, NGF, BDNF, EGF＋NGF and EGF＋BDNF groups. Immunocytochemistry for NSE was performed a week later to count the positive cells. Results The cultured monoclonal cells expressed nestin. The expression of NSE and GFAP were observed one week after induced differentiation. The percentage of NSE positive cells differentiated from NSCs in EGF＋NGF and EGF＋BDNF groups are much higher than those in other groups with the highest in EGF＋BDNF group. Conclusion The ability of BDNF on promoting differentiation of adult rat hippocampus NSCs into neurons is higher than NGF in the media containing EGF.

关 键 词：神经干细胞 海马 表皮生长因子 神经生长因子 脑源性神经生长因子 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]