大鼠骨髓间充质干细胞培养上清液对RSC96细胞生物学功能的影响

摘要

目的:探讨大鼠骨髓间充质干细胞(BMSC)培养上清液对大鼠RSC96细胞生物学功能的影响.方法:体外培养、纯化SD大鼠BMSC并进行鉴定;收集第3代BMSC培养上清液(BMSC-CM);采用MTT法检测BMSC-CM对RSC96细胞增殖的影响;平板克隆实验分析BMSC-CM对RSC96单个细胞增殖的影响;划痕实验和TranswellTM小室实验分析BMSC-CM对RSC96细胞迁移的作用,Western blot法检测BMSC-CM对RSC96细胞Bax和Bcl-2蛋白表达的改变.结果:SD大鼠第3代BMSC形态呈长梭形,旋涡样生长;成骨染色为阳性结果;成脂诱导后有脂滴出现;BMSC-CM作用后抑制RSC96细胞的增殖和迁移能力,且作用后的RSC96细胞Bax蛋白水平升高,Bcl-2蛋白表达降低.结论:BMSC-CM促进RSC96细胞凋亡,抑制其增殖和迁移.%Objective:To study the effect of culture supernatant of rat bone marrow mesenchymal stem cells (BMSCs) on biological characteristics of RSC96 Schwann cells.Methods: BMSCs of Sprague Dawley rat were cultured and purified in vitro.The BMSCs culture supernatant medium (BMSC-CM) was collected from the third generation of BMSCs.Using MTT method to detect proliferation influence of BMSC-CM on RSC96 cells,and using plate cloning assay to analysis a single cell proliferation effect.The migration analysis of BMSC-CM on RSC96 cells were detected by scratch and TranswellTM chamber assay,Western blot tested Bax and Bcl-2 proteins changes in RSC96 cells.Results: The third generation of BMSCs was long spindle-shaped and vortex-like;the result of Osteogenesis staining was positive;lipid droplets appeared after adipogenesis induction.BMSC-CM inhibited the proliferation and migration of RSC96 cells.Meanwhile,Bcl-2 protein decreased and Bax protein increased in RSC96 cells after BMSC-CM treatment.Conclusion: BMSC-CM can promote apoptosis and inhibit the proliferation and migration of RSC96 cells.