碱性成纤维细胞生长因子预诱导对骨髓基质干细胞向多巴胺能神经元分化的影响.doc

碱性成纤维细胞生长因子预诱导对骨髓基质干细胞向多巴胺能神经元分化的影响 碱性成纤维细胞生长因子预诱导对骨髓基质干细胞向多巴胺能神经元分化的影响 作者：陈欢意，牛平，赵帅，杜迎春 作者单位：110016 沈阳军区总医院神经内科 【摘要】 目的 探讨碱性成纤维细胞生长因子（bFGF）预诱导对骨髓基质干细胞（MSCs）向多巴胺（DA）能神经元分化的影响。方法 取雄性Wistar大鼠股骨和胫骨骨髓，进行MSCs的体外培养、传代扩增及纯化。bFGF预诱导24 h后，依据加入的神经营养因子不同分为单唾液酸四己糖神经节苷脂（GM1）组、胶质源性神经营养因子（GDNF）组和GDNF+GM1组,以及对照组。倒置显微镜下观察细胞形态变化，分别在预诱导第3 d、7 d进行神经元特异性烯醇化酶（NSE）、神经胶质酸性蛋白（GFAP）、酪氨酸羟化酶（TH）免疫细胞化学检测。计数NSE和TH阳性细胞数，并计算阳性细胞百分比。结果 对照组见少量NSE阳性细胞。实验组于诱导第3 d、7 d见较多数量的NSE、TH阳性细胞，GFAP阴性。bFGF预诱导各组中GDNF+GM1组NSE、TH阳性细胞率最高，GDNF组次之，GM1组最低，组间比较差异有统计学意义(均P0.01)。结论 bFGF预诱导不仅可明显促进GDNF、GM1诱导MSCs向神经元样细胞分化，表达神经元细胞标志物mdash;mdash;NSE；还可促进MSCs向DA能神经元分化，表达DA能神经元标志物mdash;mdash;TH。 【关键词】 碱性成纤维细胞生长因子；骨髓基质干细胞；多巴胺能神经元 　　Effects of basic fibroblast growth factor preinducing on marrow stroma cells which were induced into dopaminergic neurons in vitro CHEN Huanyi, NIU Ping, ZHAO Shuai, et al.Department of Neurology, General Hospital of Shenyang Military Command, Shenyang 110016, China Abstract：Objective To explore the effects of basic fibroblast growth factor (bFGF) preinducing on marrow stroma cells (MSCs) which were induced into dopaminergic neurons.Methods The rat MSCs were isolated primarily from the femurs and tibias of the Wistar rats. MSCs were cultured, proliferated and purified by passage culture. After being induced by bFGF, cultuered MSCs were divided into experimental groups (GM1 group, GDNF group and GDNF+ GM1 group) and control group. The surface markers of the differentiated neuron, such as neurone specific enolase (NSE), glial fibrillary acidic protein (GFAP) and TH were detected by immunocytochemistry after MSCs were cultured in induction media for 3 days and 7 days.Results In control group, the NSE expression of MSCs was very low. Many NSEpositive cells and THpositive cells were found in the experimental groups at 3 or 7 days after induction. The percentage of NSEpositive and THpositive cells of GDNF+GM1 group was significantly more than the other experim