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.PDF

马永宾等摇 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摇 第9期 ·1291· doi:10.3969/ j.issn.1000鄄484X.2017.09.003 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淤 于 马永宾摇 葛锁华摇 汪雪峰 摇 苏建华摇 (江苏大学附属金坛医院,金坛 213200) 摇 摇 中图分类号摇 R392鄄33摇 Q255摇 Q954郾67+3摇 摇 文献标志码摇 A摇 摇 文章编号摇 1000鄄484X(2017)09鄄1291鄄05 [摘摇 要]摇 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鄄CM);采用MTT法检测BMSC鄄CM对RSC96细胞增 TM 殖的影响;平板克隆实验分析BMSC鄄CM对RSC96 单个细胞增殖的影响;划痕实验和Transwell 小室实验分析 BMSC鄄CM对 RSC96细胞迁移的作用,Western blot法检测 BMSC鄄CM对 RSC96 细胞 Bax 和Bcl鄄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鄄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鄄2蛋白表达降低。 结论:BMSC鄄CM促进RSC96 细胞凋亡,抑制其增 殖和迁移。 [关键词]摇 骨髓间充质干细胞;RSC96细胞;增殖;迁移 Effect of culture supernatant of rat bone mesenchymal stem cells on biological characteristics of RSC96 cells MA Yong鄄Bin, GE Suo鄄Hua, WANG Xue鄄Feng,SU Jian鄄Hua郾Jintan Hospital, Jiangsu University, Jintan 213200,China [Abstract]摇 Objective:To study the effect of culture supernatant of rat bone marrow mesenchymal stem cells (BMSCs) on biological characteristics of RSC96 Schwann cells郾Methods:BMSCs of Sprague Dawley rat were cultured and purified in vitro郾The BMSCs culture supernatantmedium (BMSC鄄CM)wascollectedfromthethirdgenerationof BMSCs郾UsingMTTmethodtodetectprolif鄄 eration influence of BMSC鄄CM on RSC96 cells,andusingplate cloning assay to analysisa single cell proliferation effect郾Themigration TM analysis of BMSC鄄CM on RSC96 cells were detected by scratch and Transwell chamber assay,Western blot tested Bax and Bcl鄄2 proteins changes in RSC96 cells郾Results: The third generation of BMSCs was long spindle鄄shaped and vortex鄄like;the result of Osteogenesis staining was positive;lipid droplets appeared after adipogenesis i