软骨源性形态发生蛋白-2诱导骨髓间充质干细胞表达软骨细胞表型的实验研究

发布时间：2018-11-02 17:18

【摘要】： 目的 研究软骨源性形态发生蛋白-2(CDMP-2)对大鼠骨髓间充质干细胞(BMSCs)向软骨分化的作用。 方法 体外培养大鼠骨髓间充质干细胞(BMSCs),贴壁细胞传代,取第3代细胞,对照组(A组)以L-DMEM无血清培养液培养,实验组(B、C、D组)以L-DMEM无血清培养液培养,分别加入CDMP-2(100 ng/ml)、TGF-β1(10 ng/ml)和CDMP-2(100ng/ml)+TGF-β1(10 ng/ml),倒置相差显微镜观察细胞形态,14天后终止培养,行甲苯胺蓝染色,免疫组织化学染色和Western blot检测软骨特异性Ⅱ型胶原的表达。 结果 诱导14天后可见BMSCs形态由梭形向软骨细胞的多边形转变。甲苯胺蓝染色示糖胺聚糖(GAGs)均匀分布于基质中,免疫组织化学染色和Western blot示实验组Ⅱ型胶原表达阳性,且CDMP-2+TGF-β1组细胞表达Ⅱ型胶原水平高于单独给予CDMP-2、TGF-β1组(P＜0.05);对照组未见阳性表达。 结论 CDMP-2能够在体外定向诱导大鼠骨髓间充质干细胞(BMSCs)向软骨方向分化;与TGF-β1相比,CDMP-2诱导能力更强;CDMP-2与TGF-β1在诱导BMSCs向软骨分化时具有协同作用,两者合用成软骨的效应最大。
[Abstract]:Objective to study the effect of chondrogenic morphogenetic protein-2 (CDMP-2) on the differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into cartilage. Methods (BMSCs), adherent cells of rat bone marrow mesenchymal stem cells were cultured in vitro. The third passage cells were obtained. The control group (A group) was cultured in L-DMEM serum-free medium. In group D, CDMP-2 (100 ng/ml), TGF- 尾 1 (10 ng/ml) and CDMP-2 (100ng/ml) TGF- 尾 1 (10 ng/ml) were added to L-DMEM serum-free medium to observe cell morphology. After 14 days, the culture was terminated, toluidine blue staining, immunohistochemical staining and Western blot were used to detect the expression of cartilage specific type 鈪，

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