骨形态反应蛋白-2对骨髓基质干细胞与软骨细胞共培养向软骨细胞分化的影响

Effects of BMP-2 on promote of chondrogenesis in bone marrow stromal stem cells and chondrocytes co-culture

目的研究细胞因子骨形态反应蛋白-2(BMP-2)与软骨细胞共同诱导骨髓基质干细胞(BMSCs)向软骨细胞分化是否有协同作用,并检测BMP-2对BMSCs和软骨细胞共培养诱导分化为软骨细胞的最佳浓度。方法将体外分别培养扩增的兔BMSCs和软骨细胞按相同的比例(7∶3)混合后,均以5.0×107.mL-1的细胞浓度接种混合培养,根据不同的BMP-2浓度分为以下几组,5、10、20、30、40、50 ng/mL组,以单独培养的软骨细胞作为阳性对照组,以单独培养的BMSCs作为阴性对照组,以混合培养后不加BMP-2作为0 ng/mL组。通过MTT、Ⅱ型胶原蛋白表达以及糖胺聚糖蛋白表达的检测,确定BMP-2与软骨细胞共同作用于BMSCs是否有协同作用及促进BMSCs向软骨细胞转化的最佳BMP-2浓度。结果单独培养BMSCs的阴性对照组向软骨细胞分化的倾向不明显,而与软骨细胞混合培养的0 ng/mL组软骨细胞增多,添加BMP-2的5、10、20、30、40、50 ng/mL组向软骨细胞分化更加明显。结论 BMP-2与软骨细胞共同作用于BMSCs对于其向软骨细胞分化有协同作用,BMP-2的最佳作用浓度为20 ng/mL。

Objective To evaluate the efficacies of different concentration of bone morphogenetic protein 2 （ BMP - 2） on chondrogenesis of bone marrow stromal cells （BMSCs） co - cuhured with chondrocytes, and to search the optimal concentration of BMP - 2. Methods Rabbit BMSCs and chondrocytes were isolated, cultured and expanded, and co - cultured in the ratio ofT： 3 （BMSCs： chondrocytes）. BMP -2 of 0, 5, 10, 20, 30, 40 and 50 ng/mL were added in- to co - cultured cells. Chondrocytes and BMSCs cultured alone were taken as positive and negative controls, respectively. The OD of MTT, and the expressions of collagen type 11 and the GAG proteins were measured. Results Chondrocytes induced chondrogenic differentiation of BMSCs in vitro. BMP - 2 was proved to provide improvement of BMSC chondrogen- esis. Conclusion BMP - 2 promotes chondrogenesis of BMSCs co - cultured with chondrocytes, with optimal concentra- tion of 20 ng/mL.