不同鼠龄生后大鼠海马神经干细胞的传代和分化.pdfVIP

解剖科学进展 Progress of Anatomical Sciences 2∞8， 14(1):28-31 不同鼠龄生后大鼠海马神经干细胞的传代和分化 王振宇\季丽莉，佟雷，唐源远，赵久红 (中国医科大学基础医学院人体解剖学教研室，辽宁沈阳 11仪阳) [摘要]目的研究不同鼠龄大鼠海马神经干细胞 (NSCs) 的传代增殖和分化情况。方法 将生后SD大鼠 分为3d 、 10d 、 20d 3组，应用膜酶消化法将海马组织制成单细胞悬液，用含碱性成纤维生长因子 (bFGF) 、表 皮生长因子 (EGF) 、 B27的无血清细胞培养技术进行体外培养，单克隆培养后通过Nesti队 NSE、 GFAP免疫细 胞化学染色鉴定神经干细胞、神经元、星形胶质细胞;细胞传3代后，在各组培养基中加入终浓度为10%的血清 诱导分化， 1 周后进行NSE免疫细胞化学染色，计数阳性细胞比例，进行统计学分析。结果 3组SD大鼠海马组织 细胞，单克隆培养后表达Nestin ，诱导分化后分别表达NSE和GFAP。生后3d大鼠神经干细胞在传1-6代时每代均 快于生后lOd和20d大鼠，传6代后， 3组细胞传代时间间隔几近一致;生后3d组大鼠海马神经干细胞分化为神经元 的比例较其它2组高 (P0.05 )。结论生后3d大鼠神经干细胞增殖速度和分化为神经元的数量都高于1 0d和 20d大鼠。 [关键词]神经干细胞;大鼠;海马;细胞培养;免疫细胞化学法 [中图分类号] R338.2飞 [文献标识码] A [文章编号] 1 ∞6-2947 (2008) 01-∞28-04 Passage and D Ïfferentiation of N eural Stem Cells from the Hippocampus of the D Ïfferent N eonate Rats W ANG Zhen-yu，jI ιi-li，TONG Lei ,TANG Yuan-yuan ,ZHAO Jiu-hong (Department of Anatomy , China Medical University , Liaoning Shenyang 11创)()I China) [ Abstract] 0电jective To investigate the proliferation and differentiation of neural stem cells (NSCs) from hippocampus of neonate rats wi出 different ages. Methods By using trypsin digestion , neural stem cells were isolated from hippocampus of 3-d呵， 10-day and 20-day rats. The medium of serum-free but wi出 basic fibroblast growth factor (bFG F), epidennal growth factor (EGF) and B27 was used to culture NSCs. Immunocytochemistry for nestin was used to identify monoclone NSCs , for glial fibrillary acidic protein (GFAP) and neurone specific enolase (NSE) to identify astrocytes and neurons respectively one week after NSCs were differentiated. The percentage of the NSE positive cells wa