小鼠骨髓基质细胞诱导分化为神经细胞的实验研究

Experimental study of the mouse bone marrow stromal cells differentiated into neural-like cells

目的 探讨小鼠骨髓基质细胞体外诱导及向神经细胞方向分化。方法 从小鼠骨髓中分离培养骨髓基质细胞 (bonemarrowstromalcells ,BMSC) ,应用碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor ,bFGF) ,维甲酸 (retinoicacid ,RA) ,二甲亚砜和神经生长因子 (nervegrowthfactor,NGF)对细胞诱导分化 ,72h后对细胞表达神经微丝蛋白 (NF 2 0 0 ) ,胶质原性纤维酸性蛋白 (GFAP) ,纤维结合素 (fibronectin)的结果进行荧光和组化检测。结果 诱导 72h后细胞形态类似神经细胞改变 ,免疫组化显示约 60 %细胞分化为NF 2 0 0阳性细胞 ,约 5 0 %细胞分化为GFAP阳性细胞 ,同时细胞fibronectin表达明显降低 ,与对照组比较相差显著。结论 维甲酸 ,神经生长因子 ,二甲亚砜 。

Objective To observe the growth and differentiation of the cultured mouse bone marrow stromal cells in vitro into neural like cells. Methods Bone marrow stromal cells (BMSCs) isolated from mice were cultured according to the routine method, and induced to differentiate by basic fibroblast growth factor b (bFGF), retinoic acid (RA), nerve growth factor (NGF), and dimethylsulfoxide (DMSO). Expressions of neural microfilament protein (NF 200), glial fibrillary acidic protein (GFAP), and fibronectin at 72 h after inductin were determined by immunoflurenscence and immunocytochemicalmethod. Results At 72 h after inductin, neural like changes were found in BMSCs. Immunohistochemical stain showed that 60% BMSCs differentiated into NF 200 positive cells, and about 50% BMSCs into GFAP positive cells. Fibronectin expression was significantly lower as compared with that in the control group. Conclusion Combined action of RA, bFGF, NGF, and DMSO can induce BMSCs to differentiate into neural like cells in vitro .