兔骨髓基质干细胞体外诱导分化为雪旺样细胞的实验研究

发布时间：2018-03-22 06:00

本文选题：骨髓基质干细胞　切入点：细胞分化　出处：《广西医科大学》2006年硕士论文　论文类型：学位论文

【摘要】： 目的:探讨体外分离及扩增纯化兔骨髓基质干细胞的实验方法,并研究其生物学特性;以不同诱导液在体外对BMSCs诱导分化为雪旺样细胞,探讨以BMSCs作为周围神经组织工程种子细胞的可行性。方法:采用Percoll(密度为1.073g/ml)分离液和贴壁筛选法,从兔股骨中分离BMSCs进行纯化、培养;原代细胞按1: 2比例进行传代培养;在DMEM-LG + 10%FBS中培养,用β-ME、b-FGF和黄芪等为诱导剂对第1, 3, 5代骨髓基质干细胞进行定向诱导分化,倒置像差显微镜观察细胞形态学化,免疫细胞化学鉴定S-100蛋白和GFAP的表达率。结果:原代及传代培养显示,兔BMSCs具有活跃的增殖能力,第2, 3, 5代细胞的生长曲基本相同;诱导后的兔骨髓基质干细胞具有类似雪旺细胞的形态,免疫细胞化学鉴定显示S-100蛋白和GFAP的表达率(76±6.7)%、(25±5.45)%。结论:应用本实验方法培养兔髓基质干细胞,体外生长稳定、增殖速度快、可连续传代;采用β-ME、b-FGF和黄芪等为诱导剂,可使兔骨髓基质干细胞在体外向雪旺样细胞分化。
[Abstract]:Objective: to investigate the methods of isolating and purifying rabbit bone marrow stromal cells in vitro and their biological characteristics, and to differentiate Schwann like cells into Schwann like cells induced by different inducers in vitro. To explore the feasibility of using BMSCs as seed cells for peripheral nerve tissue engineering. Methods: BMSCs isolated from rabbit femur was purified and cultured by Percollus (density 1.073 g / ml) and adherent screening method, primary cells were subcultured in 1: 2 ratio, and cultured in DMEM-LG 10s. Bone marrow stromal stem cells (BMSCs) of the first, third and fifth passages were induced to differentiate by 尾 -ME-b-FGF and astragalus membranaceus. The morphologic changes of bone marrow stromal cells were observed by inverted aberration microscope. The expression rates of S-100 protein and GFAP were identified by immunocytochemistry. Results: primary culture and passage culture showed that rabbit BMSCs had active proliferative ability, and the growth curve of the 2nd, 3rd and 5th passage cells was basically the same, and the induced rabbit bone marrow stromal stem cells had the morphology similar to Schwann cells, and the growth curve of rabbit bone marrow stromal cells was similar to that of Schwann cells. The expression rate of S-100 protein and GFAP was 76 卤6.7% and 25 卤5.45% by immunocytochemistry. Conclusion: rabbit marrow stromal stem cells can be cultured in vitro with stable growth, rapid proliferation and continuous passage, and 尾 -ME-b-FGF and Astragalus membranaceus can induce the differentiation of rabbit bone marrow stromal cells into Schwan-like cells in vitro.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329.2

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