文章摘要
吕 立,史德志,陈 功,张 春,顾 斌,胡卫星.顺序性化学诱导促大鼠骨髓间充质干细胞向多巴胺能神经元分化[J].南京医科大学学报,2010,(5):621~625
顺序性化学诱导促大鼠骨髓间充质干细胞向多巴胺能神经元分化
Chemically defined sequential culture media for dopaminergic neuron-like cells from rat bone mesenchymal stem cells
投稿时间:2009-11-04  
DOI:10.7655
中文关键词: 顺序性化学诱导  骨髓间充质干细胞  分化  多巴胺神经元
英文关键词: chemically defined sequential culture  bone marrow mesenchymal stem cells  differentiation  dopaminergic neurons
基金项目:江苏省科技厅基础研究计划(自然科学基金)(BK2007257)
作者单位
吕 立 南京医科大学第一附属医院神经外科,江苏 南京 210029 
史德志  
陈 功  
张 春  
顾 斌  
胡卫星  
摘要点击次数: 1233
全文下载次数: 170
中文摘要:
      目的:研究大鼠骨髓间充质干细胞(BMSCs)在顺序性化学诱导培养基诱导下向多巴胺(DA) 能神经元细胞分化的能力?方法:通过密度梯度离心法获取大鼠骨髓中的单个核细胞,贴壁培养纯化BMSCs?以碱性成纤维细胞生长因子(bFGF)?表皮生长因子(EGF)?神经条件培养基(NCM)?音速波状蛋白(SHH)和成纤维细胞生长因子8(FGF8)联合对第4代BMSCs进行诱导?RT-PCR?Western blot和细胞免疫荧光染色检测DA能神经元分化过程中的标志物酪氨酸羟化酶(TH)?多巴胺转运蛋白(DAT)的表达以及转录因子Nurr1?TUJ1?AADC 和VAMT的表达?结果:诱导2周后,RT-PCR结果显示Nurr1?AADC?VAMT?DAT和TH的mRNA均有表达;Western blot显示TH和TUJ1蛋白均有表达,细胞免疫荧光染色结果表明诱导2周后TH阳性细胞占(41.18±3.24) %?结论:按一定顺序给予化学诱导剂可以在体外诱导BMSCs向DA能神经元分化,并具有DA能神经元的功能特征,是临床用于治疗帕金森病这一类神经精神性疾病的理想细胞来源?
英文摘要:
      Objective:To explore the differentiation of rat bone mesenchymal stem cells to dopaminergic neurons by chemically defined sequential culture media in vitro. Methods:The rat bone mesenchymal stem cells were isolated from rat marrow by density gradient centrifugation and the fourth generation was induced into dopaminergic neurons by treated with bFGF,EGF,NCM,SHH and FGF8. Dopamine associated proteins and genes in the treated cells were examined by Western blot,immumofluore-scence and RT-PCR. Results:After 2 weeks of stimulation,the cells expressed the characteristics of dopaminergic neurons. RT-PCR discovered the expression of mRNA of DAT,Nurr1,AADC,VAMT and TH. Immumofluorescence staining indicated that the ratio of TH positive neural cells was significantly increased after induction for 2 weeks(41.18±3.24) %. Conclusion:Chemically defined sequential culture media can induce differentiation of BMSCs into DA neurons in vitro,and the expression of mature neurons characteristics of the transdifferentiated cell. BMSCs might be a suitable and available source for the in vitro derivation of DA neurons and cell transplantation therapy on some central neural system diseases such as Parkinson’s disease.
查看全文   查看/发表评论  下载PDF阅读器
关闭