發(fā)布時(shí)間：2018-11-02 17:18

【摘要】： 目的 研究軟骨源性形態(tài)發(fā)生蛋白-2(CDMP-2)對(duì)大鼠骨髓間充質(zhì)干細(xì)胞(BMSCs)向軟骨分化的作用。 方法 體外培養(yǎng)大鼠骨髓間充質(zhì)干細(xì)胞(BMSCs),貼壁細(xì)胞傳代,取第3代細(xì)胞,對(duì)照組(A組)以L-DMEM無血清培養(yǎng)液培養(yǎng),實(shí)驗(yàn)組(B、C、D組)以L-DMEM無血清培養(yǎng)液培養(yǎng),分別加入CDMP-2(100 ng/ml)、TGF-β1(10 ng/ml)和CDMP-2(100ng/ml)+TGF-β1(10 ng/ml),倒置相差顯微鏡觀察細(xì)胞形態(tài),14天后終止培養(yǎng),行甲苯胺藍(lán)染色,免疫組織化學(xué)染色和Western blot檢測(cè)軟骨特異性Ⅱ型膠原的表達(dá)。 結(jié)果 誘導(dǎo)14天后可見BMSCs形態(tài)由梭形向軟骨細(xì)胞的多邊形轉(zhuǎn)變。甲苯胺藍(lán)染色示糖胺聚糖(GAGs)均勻分布于基質(zhì)中,免疫組織化學(xué)染色和Western blot示實(shí)驗(yàn)組Ⅱ型膠原表達(dá)陽性,且CDMP-2+TGF-β1組細(xì)胞表達(dá)Ⅱ型膠原水平高于單獨(dú)給予CDMP-2、TGF-β1組(P＜0.05);對(duì)照組未見陽性表達(dá)。 結(jié)論 CDMP-2能夠在體外定向誘導(dǎo)大鼠骨髓間充質(zhì)干細(xì)胞(BMSCs)向軟骨方向分化;與TGF-β1相比,CDMP-2誘導(dǎo)能力更強(qiáng);CDMP-2與TGF-β1在誘導(dǎo)BMSCs向軟骨分化時(shí)具有協(xié)同作用,兩者合用成軟骨的效應(yīng)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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