兔骨髓间充质干细胞体外诱导分化为神经样细胞的实验研究

Study on neurotrophic agent inducing the differentiation of rabbit bone marrow mesenchymal stem cells into neuron like cells

目的动态观察体外神经营养剂联合碱性成纤维细胞生长因子(bFGF)定向诱导骨髓间充质干细胞(BMSCs)分化为神经样细胞的效果。方法将BMSCs分离体外扩增培养后,分别加入神经营养剂金路捷、三磷酸腺苷(ATP)、神经节苷脂(GM1)+bFGF诱导其分化。分别于6、24、48、72、96h后光镜下观察细胞形态变化并通过免疫组织化学测定特异性神经元稀醇化酶(NSE)、神经巢蛋白(Nestin)、胶质纤维酸性蛋白(GFAP)的表达情况。结果神经营养剂联合bFGF诱导6h后出现胞体收缩,Nestin表达阳性,24h后伸出突起,有不同程度NSE、GFAP表达,3d后达高峰。结论神经营养剂联合bFGF能提高诱导骨髓间充质干细胞分化为神经样细胞的诱导率。

Objective To observe the effects of the neurotrophic agent combined with basic fibroblast growth factor （bFGF） on inducing rabbit bone marrow mesenchymal stem cells （BMSCs） to differentiate into neuron like cells. Methods BMSCs were separated and cultured and expanded in vitro, then BMSCs were induced with mouse nerve growth factor for injection and adenosine triphosphate（ATP） and gangliosides（ GM1） ＋ bFGF. Neuron morphology was observed and the expression of neurofilament protein （NSE） ＋ , Nestin ＋ , glia fibrillary protein （GFAP） ＋cells were detected with immunocytochemistry at 6, 12, 24, 48, 72 h. Results The cells body contracted after 6 h, and the nestin were positive. After 24 h there were neurites grew from cells, the expression of NSE and GFAP were positive. Three days later the positive cells exhibited value peak. Conclusion Combination of neurotrophic agent with hFGF synergistically promotes the transformation of BMSC and differentiation into neuron like cells.