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目的探讨骨髓间充质干细胞(Bone mesenchymal stem cells,BMSCs)对大鼠脑胶质瘤C6细胞系分化的影响及其相关机制。方法用Transwell小室共培养BMSCs与C6细胞,采用细胞计数法检测C6细胞增殖水平;流式细胞术检测C6细胞的细胞周期;免疫荧光与免疫组化法分别检测波形蛋白(vimentin)和胶质纤维酸性蛋白(Glia lfibrillary acidic protein,GFAP)的表达;生化法检测谷氨酰胺合成酶(Glutamine synthetase,GS)的活性。结果 BMSCs对C6细胞的增殖具有抑制作用;与BMSCs进行双层培养72h的C6细胞出现G0/G1期细胞周期阻滞,双层培养组的C6细胞GFAP表达明显增强,同时vimentin表达减弱,并且这两种分化标志物在细胞内的分布及排列发生了改变;双层培养组C6细胞内GS活性升高。结论 BMSCs能诱导C6细胞向成熟星形胶质细胞分化,可能与BMSCs分泌的某些细胞因子有关。
Objective To investigate the effect of Bone marrow mesenchymal stem cells (BMSCs) on the differentiation of C6 glioma cells in rats and its related mechanism. Methods BMSCs and C6 cells were co-cultured in Transwell chambers. The proliferation of C6 cells was detected by cell counting. The cell cycle of C6 cells was detected by flow cytometry. The expressions of vimentin and glial fibrinogen were detected by immunofluorescence and immunohistochemistry. The expression of Glia lfibrillary acidic protein (GFAP) was detected by biochemical method. The activity of Glutamine synthetase (GS) was detected by biochemical method. Results BMSCs had an inhibitory effect on the proliferation of C6 cells. C6 cells cultured in BMSCs for 72h exhibited G0 / G1 phase cell cycle arrest, while GFAP expression in C6 cells in BMSCs was significantly increased and vimentin expression was weakened at the same time The distribution and arrangement of the two differentiating markers in the cells changed; the activity of GS in C6 cells in double-layer culture group increased. Conclusion BMSCs can induce the differentiation of C6 cells into mature astrocytes, which may be related to some cytokines secreted by BMSCs.