大鼠骨髓源神经干细胞的分离培养及鉴定  

Isolation,Culture and Characterization of Bone Marrow Mesenchymal Stem Cells-derived Neural Stem Cells of Wistar Rats

作　　者：陈晓娟[1] 肖宗宇[2] 潘琪[3] 侯倩[1] 才鼎[1] 周有婷 

机构地区：[1]青海省人民医院,青海西宁810007 [2]青海大学附属医院 [3]海南医学院附属医院

出　　处：《中国医学创新》2015年第28期18-22,共5页Medical Innovation of China

基　　金：青海省科技创新能力促进计划项目(2015-ZJ-943Q);青海大学中青年科研基金项目(2014-QYY-6)

摘　　要：目的:对Wistar大鼠骨髓源神经干细胞进行分离、培养和鉴定,观察其生长方式和分化特征。方法:运用无血清培养基对Wistar大鼠骨髓基质细胞进行培养,对分离获得的悬浮生长的神经球采用免疫组织化学法检测CD133和Nestin表达情况。用血清诱导其分化,分化7 d后,采用免疫荧光细胞化学染色方法检测分化后GFAP、Map2、β-tubulinⅢ、Galc的表达情况。结果:大鼠骨髓基质细胞在无血清培养基中,呈悬浮状态生长,形成细胞球,经免疫荧光检测,细胞球呈CD133和Nestin阳性。将细胞球转入含血清培养基后,转为贴壁生长,经免疫荧光检测大部分已分化细胞呈GFAP阳性,少部分细胞呈MAP-2、β-tubulinⅢ及Galc阳性。结论:采用含bFGF、EGF的无血清培养基可培养出呈球状聚集生长、具多向分化潜能的大鼠骨髓源神经干细胞。Objective： To isolate, culture and identify the bone marrow mesenchymal stem cells-derived neural stem cells （ BMSCs-NSCs ） in order to observe their growth pattern and differentiation features.Method： Bone marrow mesenchymal stem cells of the Wistar rats were cultured in serum-free medium.Floating neural spheres were formed in serum-free medium, the expression of CD133 and Nestin in the neural spheres was tested by immunohistochemistry.Then floating neural spheres were transferred into serum containing medium for induction differentiation, the expression of GFAP, Map2, 13 -tubulin Ⅲ and Galc were detected by immunocytochemistry 7 days later.Result： Bone marrow mesenchymal stem cells lived in serum-free medium ＇floating＇ and it formed neural sphere.CD133 and Nestin were positive in neural sphere cells according to the results of immunohistochemistry.When these neural sphere ceils were transferred into serum-containing medium, they grew adhering to the wall.GFAPMAP-2, β -tubulin m and Galc were positive in the cells differentiated from neural spheres.Conclusion： Bone marrow mesenchymal stem cells-derived neural stem cells which growing like a ball and having muhipotential differentiation are successfully cultured in serum-free medium with bFGF and EGF.

关 键 词：骨髓基质细胞 神经干细胞 WISTAR大鼠 CD133 NESTIN 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]