定向诱导骨髓基质干细胞向神经元样细胞分化的实验研究

An experimental study on targeted induction of human bone mesenchymal stem cells to neuron-like cells

目的建立一套系统的体外分离、培养及诱导人骨髓基质干细胞（BMSCs）分化为神经元样细胞的方法。方法采用人淋巴细胞分离液从骨髓中分离出BMSCs，体外培养、扩增后，加以复合神经诱导剂进行诱导，诱导期间观察细胞形态的变化，并通过免疫细胞化学鉴定诱导细胞表面标志物的表达情况。结果人BMSCs在体外分离、培养、扩增后，经复合神经诱导剂诱导48h后，部分细胞出现胞体收缩和突起伸出，呈现神经元细胞样改变。免疫细胞化学染色显示巢蛋白或鼠抗人神经丝单克隆抗体阳性，兔抗人胶质纤维酸性蛋白阴性。结论本实验成功培养并诱导人BMSCs分化为神经元样细胞，并建立了一套系统的实验方法。

Objective To establish a systematic method of isolating, culturing and inducing human bone mesenchymal stem cells （hBMSCs） to neuron-like cells in vitro. Methods The hMSCs were isolated from bone marrow with the lymphocyte separating medium, cultured and expanded in vitro, and induced by compound neuro-revulsants. The morphologic changes of cells were observed, and the expressions of surface markers of induced cells were identified by immunocytochemistry during induction period. Results hBMSCs could be separated, cultured and expanded in vitro. After induction by combined neuro-revulsants for 48 hours, neuronal marks, such as cellular shrinkage and neurite growth, were observed in some of the cells. The immunochemical staining revealed nestin or NF （neurofilament） positive, and GFAP （glial fibrillary acidic protein） negative. Conclusion The hMSCs can be cultured and induced into neuron-like cells in experiment.