骨髓间充质干细胞分化神经元后多巴胺D2受体的表达

Expression of dopamine receptor D2 in neurons derived from mesenchymal stem cells

目的:检测骨髓间充质干细胞(mesenchymal stem cells,MSCs)分化为神经元后多巴胺D2受体(dopamine receptor D2,DRD2)的表达,探讨体外分化神经元的基本功能。方法:采用密度梯度离心法和贴壁培养的方法分离纯化MSCs,流式细胞仪检测MSCs表面标志,DMSO/BHA/FSK作为诱导剂来诱导分化;硝酸银染色法检测神经纤维丝,RT-PCR和免疫细胞化学法检测NSE、DRD2 mRNA及蛋白的表达情况。结果:分离纯化的MSCs经流式细胞仪检测显示CD34、CD45表达阴性;间充质干细胞相对特异性标志CD44强阳性表达;加入诱导剂后,MSCs形成多极神经元的形态。银染色法显示神经纤维丝阳性表达,RT-PCR显示诱导后细胞表达NSE、DRD2 mRNA,免疫组化显示NSE、DRD2蛋白表达阳性。结论:MSCs分化为神经元后可以表达神经受体DRD2。

Objective： To investigate the expression of dopamine receptor D2 （DRD2） in the neuron-like cells derived from mesenchymal stem cells （MSCs）. Methods： MSCs were separated from adult bone marrow with density gradient centrifugation and adherent culture, induced and differentiated into neuron-like cells with DMSO/BHA/FSK. Expression of cell surface marker on MSCs was detected by flow cytometer, neurofibra by silver staining, protein of DRD2 and NSE by immunocytochemistry, and mRNA of DRD2 and NSE by RT-PCR. Results： Flow cytometry showed that CD44 on MSCs was positive,while CD34 and CD45 were negative; silver staining showed that MSCs rapidly presented the morphology of muhipolar neurons and neurofibra 1 d after DMSO/BHA/FSK induction. RT-PCR showed that mRNA of NSE and DRD2 was positive. Immunocytochemistry staining indicated that protein of NSE and DRD2 were positive, too. Conclusion： MSCs could be induced into neuron expressing DRD2.