大鼠骨髓间充质干细胞促进RSC96细胞凋亡并抑制其增殖和迁移
发布时间：2018-10-15 10:50
【摘要】：目的探讨大鼠骨髓间充质干细胞(BMSC)对大鼠RSC96施万细胞增殖及迁移的影响。方法全骨髓细胞贴壁法分离并提取SD大鼠BMSC,倒置显微镜观察第3代BMSC形态;流式细胞术检测细胞表面CD19、CD34、CD73、CD105;第3代BMSC经成脂成骨诱导液分别诱导3周和2周后,分别应用油红O染色和碱性磷酸酶检测BMSC的多向分化能力。应用24 mm共培养板,将第3代BMSC与RSC96细胞共培养24 h,采用MTT法和平板克隆形成实验检测RSC96细胞增殖及克隆形成能力;划痕实验及TranswellTM小室实验检测RSC96细胞的迁移能力;Western blot法检测RSC96细胞Bax和Bcl-2蛋白表达水平。结果SD大鼠第3代BMSC显微镜下呈梭形、旋涡状排列,形态大小相似;BMSC表面标志CD73、CD105高表达,造血干细胞表面标志CD34和淋巴细胞表面标志CD19低表达;BMSC经成脂诱导培养3周后,细胞有大量脂滴;经成骨诱导培养2周后,碱性磷酸酶染色呈阳性;BMSC与RSC96细胞共培养后,与对照组相比,RSC96细胞的增殖活性显著降低,克隆形成能力显著减弱、细胞迁移能力降低;共培养后RSC96细胞Bax蛋白水平升高,Bcl-2蛋白表达降低。结论 BMSC能促进RSC96细胞凋亡,抑制其增殖、迁移。
[Abstract]:Objective to investigate the effect of rat bone marrow mesenchymal stem cell (BMSC) on proliferation and migration of rat RSC96 Schwann cells. Methods BMSC, of SD rats was isolated and extracted by whole bone marrow adherent method. The morphology of the third generation BMSC was observed by inverted microscope, and the third generation of BMSC on the cell surface was detected by flow cytometry after being induced by adipogenic osteoblasts for 3 weeks and 2 weeks, respectively. Oil red O staining and alkaline phosphatase were used to detect the multidirectional differentiation ability of BMSC. The third generation of BMSC and RSC96 cells were co-cultured for 24 h with 24 mm co-culture plate. The proliferation and clone forming ability of RSC96 cells were detected by MTT assay and plate clone formation assay. The migration ability of RSC96 cells was detected by scratch test and TranswellTM chamber assay. The expression of Bax and Bcl-2 in RSC96 cells was detected by; Western blot method. Results the third generation BMSC of SD rats showed spindle shape, swirl arrangement and similar size under microscope, high expression of CD73,CD105 on BMSC surface, low expression of CD34 on hematopoietic stem cell surface and CD19 on lymphocyte surface, BMSC was induced by adipogenic culture for 3 weeks. After 2 weeks of osteogenic induction, alkaline phosphatase staining was positive. After co-culture of BMSC and RSC96 cells, the proliferation activity of RSC96 cells was significantly lower than that of the control group, and the ability of clone formation was significantly decreased. The level of Bax protein increased and the expression of Bcl-2 protein decreased after co-culture. Conclusion BMSC can promote apoptosis, inhibit proliferation and migration of RSC96 cells.
【作者单位】： 江苏大学附属金坛医院;江苏大学附属医院;
【基金】：江苏省自然科学基金(BK20141295) 常州市卫生局重大科研立项(ZD201508)
【分类号】：R329.2
，
本文编号：2272313
[Abstract]:Objective to investigate the effect of rat bone marrow mesenchymal stem cell (BMSC) on proliferation and migration of rat RSC96 Schwann cells. Methods BMSC, of SD rats was isolated and extracted by whole bone marrow adherent method. The morphology of the third generation BMSC was observed by inverted microscope, and the third generation of BMSC on the cell surface was detected by flow cytometry after being induced by adipogenic osteoblasts for 3 weeks and 2 weeks, respectively. Oil red O staining and alkaline phosphatase were used to detect the multidirectional differentiation ability of BMSC. The third generation of BMSC and RSC96 cells were co-cultured for 24 h with 24 mm co-culture plate. The proliferation and clone forming ability of RSC96 cells were detected by MTT assay and plate clone formation assay. The migration ability of RSC96 cells was detected by scratch test and TranswellTM chamber assay. The expression of Bax and Bcl-2 in RSC96 cells was detected by; Western blot method. Results the third generation BMSC of SD rats showed spindle shape, swirl arrangement and similar size under microscope, high expression of CD73,CD105 on BMSC surface, low expression of CD34 on hematopoietic stem cell surface and CD19 on lymphocyte surface, BMSC was induced by adipogenic culture for 3 weeks. After 2 weeks of osteogenic induction, alkaline phosphatase staining was positive. After co-culture of BMSC and RSC96 cells, the proliferation activity of RSC96 cells was significantly lower than that of the control group, and the ability of clone formation was significantly decreased. The level of Bax protein increased and the expression of Bcl-2 protein decreased after co-culture. Conclusion BMSC can promote apoptosis, inhibit proliferation and migration of RSC96 cells.
【作者单位】： 江苏大学附属金坛医院;江苏大学附属医院;
【基金】：江苏省自然科学基金(BK20141295) 常州市卫生局重大科研立项(ZD201508)
【分类号】：R329.2
，
本文编号：2272313
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