Ma Yongbin, Ge Suohua, Zhou Dan, Zhang Huanyan, Sun Luyang, Wang Xuefeng, Su Jianhua
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2017 Feb;33(2):190-5.
To investigate the effect of rat bone marrow mesenchymal stem cells( BMSCs) on the proliferation and migration of rat RSC96 Schwann cells in vitro.
BMSCs of Sprague Dawley( SD) rats were isolated by bone marrow cell adherent method. The morphology of the third generation of BMSCs was observed by inverted microscopy. The cell surface antigen makers CD19,CD34,CD73,CD105 were detected by flow cytometry. The third-passage BMSCs differentiated into osteoblasts and adipocytes after exposed to bone-inducing and lipid-inducing media for 3 and 2 weeks,respectively,and then were identified by oil red O and alkaline phosphatase staining. The third generation of BMSCs was co-cultured with RSC96 cells for 24 hours in 24 mm co-culture plate. The cell proliferation and colony formation of RSC96 cells were determined by MTT assay and plate cloning assay. The migration of RSC96 cells was detected by scratch and TranswellTMchamber assay. Bax and Bcl-2 expression levels of RSC96 cells were detect by Western blot analysis.
The third generation of BMSCs showed fusiform,spiral arrangement with similar cell size under an inverted microscope. The CD73 and CD105 expression levels of BMSCs were high,however,the CD34 and CD19 were low. BMSCs had large lipid droplets after adipogenic induction for 3 weeks; and the alkaline phosphatase staining of BMSCs was positive after osteogenic induction for 2 weeks. Compared to the control group,co-culture of RSC96 cells and BMSCs induced a significant decrease in the proliferation,colony formation and migration of RSC96 cells. Meanwhile,Bax protein expression increased,and Bcl-2protein expression decreases in RSC96 cells co-cultured with BMSCs.
BMSCs could promote the apoptosis and inhibit the proliferation and migration of RSC96 cells.
探讨大鼠骨髓间充质干细胞（BMSCs）对大鼠RSC96雪旺细胞体外增殖和迁移的影响。
采用骨髓细胞贴壁法分离Sprague Dawley（SD）大鼠的BMSCs。用倒置显微镜观察第三代BMSCs的形态。采用流式细胞术检测细胞表面抗原标志物CD19、CD34、CD73、CD105。第三代BMSCs分别在成骨诱导培养基和脂肪诱导培养基中培养3周和2周后分化为成骨细胞和脂肪细胞，然后通过油红O和碱性磷酸酶染色进行鉴定。将第三代BMSCs与RSC96细胞在24 mm共培养板中共同培养24小时。采用MTT法和平板克隆法检测RSC96细胞的增殖和集落形成。采用划痕实验和TranswellTM小室实验检测RSC96细胞的迁移。通过蛋白质免疫印迹分析检测RSC96细胞中Bax和Bcl-2的表达水平。
倒置显微镜下第三代BMSCs呈梭形，螺旋状排列，细胞大小相近。BMSCs的CD73和CD105表达水平较高，而CD34和CD19表达水平较低。脂肪诱导3周后BMSCs有大量脂滴；成骨诱导2周后BMSCs碱性磷酸酶染色呈阳性。与对照组相比，RSC96细胞与BMSCs共培养后，RSC96细胞的增殖、集落形成和迁移均显著降低。同时，与BMSCs共培养的RSC96细胞中Bax蛋白表达增加，Bcl-2蛋白表达降低。
BMSCs可促进RSC96细胞凋亡，抑制其增殖和迁移。
