Wnt3a促进大鼠骨髓间充质干细胞向神经元样细胞分化

目的:寻找促进骨髓间充质干细胞(MSCs)向神经元样细胞分化的Wnt信号分子。方法:首先体外分离培养大鼠MSCs并传代,形态学观察和流式细胞学检测细胞表型CD44、CD9、CD34和CD45。采用bFGF分别联合Wnt3a或Wnt5a的方案诱导分化。免疫组化和RT-PCR的方法比较Wnt3a和Wnt5a对MSCs向神经元样细胞分化的影响。结果:MSCs为长梭形,CD9、CD44高表达,CD34、CD45低表达。Wnt3a诱导组的Nestin和NSE呈阳性,而GFAP无明显表达,诱导后细胞的活力良好。Wnt5a诱导组Nestin呈弱阳性表达,而NSE及GFAP阴性。RT-PCR结果显示Wnt3a诱导组Nestin在诱导前后均有表达,NSE在诱导后5d可见明显的扩增条带,10d后更加明显。GFAP在诱导10d后出现较弱的扩增条带。Wnt5a组、对照组MSCs在诱导后10dNestin有微弱表达,NSE和GFAP几乎无表达。结论:Wnt3a分子能够促进体外培养的MSCs向神经元样细胞分化。

Wnt3a induced rat bone marrow mesenchymal stem cells to neuron-like cells

Objective:To find out the molecules of the Wnt family which are involved in differentiation of rat bone marrow mesenchymal stem cells (MSCs) into neuron-like cells.Methods:The MSCs were separated,grained and cultured in vitro.The growth characters of the MSCs were observed.Meanwhile a analysis was taken for its phonotype.Wnt3a,Wnt5a contained bFGF were used to induce MSCs into neuron-like cells,and then identified them by detecting the NSE and GFAP using immunocytochemistry and RT-PCR.Results:The MSCs were fibroblast-like.CD9,CD44 were high expressed,but CD34,CD45 low expressed.Immunocytochemistry showed that Nestin,NSE were positive and the expression of GFAP were not obvious in Wnt3a group,the cells’ vigor were well after inducement.In Wnt5a group Nestin was weakly expressed and NSE,GFAP were negative.In Wnt3a group RT-PCR results revealed that Nestin were expressed whenever before or after inducement.NSE stripe were seen 5 d after differentiation,and became more obvious on 10 d.Blink GFAP stripe were seen on 10 d after inducement.In Wnt5a,control group Nestin were weak positive and NSE and GFAP were negative by RT-PCR.Conclusion:Wnt3a may plays a important role in differentiation of MSCs to be neuron-like cells.