BMP-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
发布时间：2019-03-11 11:05
【摘要】：目的 研究骨反应形态蛋白（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,并检测出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兔BMSCs和软骨细胞按相同的比例混合（7：3），，均以5.0×107/ml的细胞中浓度接种混合培养。根据不同的细胞因子BMP-2的浓度分为以下几组，5ng/ml,10ng/ml,20ng/ml,30ng/ml,40ng/ml，50ng/ml以单独培养软骨细胞作为阳性对照组，以单独培养BMSCs作为阴性对照组，以混合培养后不加BMP-2细胞因子作为0ng/ml组。加入生长因子培养24h、48h、72h后以四唑盐（MTT）比色法测细胞相对数(OD值)，然后分别计算BMP-2在不同浓度时于490nm处的OD值的平均值，并转换为细胞相对数，得出细胞因子BMP-2与软骨细胞共同作用于骨髓基质干细胞向软骨细胞分化和增殖有协同作用。然后用免疫细胞化学染色法确定骨髓基质干细胞分化后的细胞有Ⅱ型胶原蛋白表达为软骨细胞。通过对GAG蛋白定量的检测确定BMP-2与软骨细胞共同作用于骨髓基质干细胞向软骨细胞分化时的最佳细胞因子浓度。 结果 阴性对照组与0ng/ml组之间OD值的单因素比较P＜0.05，有统计学意义。这说明软骨细胞单独诱导骨髓基质干细胞能引起细胞数量的增加，而免疫组化可以证明增加的细胞能分泌II型胶原蛋白，为软骨细胞。当BMP-2的浓度在5ng/ml至50ng/ml范围内时测得的OD值与0ng/ml组的比较也均有统计学意义，因而两种因素共同作用时骨髓基质干细胞向软骨细胞分化显著提高，并以20ng/ml处理后OD值最高。 结论 细胞因子BMP-2与软骨细胞共同作用于骨髓基质干细胞对于其向软骨细胞分化有协同作用，且BMP-2与软骨细胞共同作用于骨髓基质干细胞向软骨细胞分化的最佳作用浓度为20ng/ml。
[Abstract]:Objective to investigate the synergistic effect of bone reactive morphogenetic protein (BMP-2) and chondrocytes on the differentiation of bone marrow stromal stem cells (BMSCs) into chondrocytes. The optimal concentration of BMP-2 on chondrocytes induced by co-culture of BMSCs and chondrocytes was determined. Methods the cultured rabbit BMSCs and chondrocytes were mixed in the same proportion (7:3) in vitro, and cultured in 5. 0 脳 107/ml medium. According to the concentration of different cytokine BMP-2, they were divided into the following groups: 5 ng / ml, 10 ng / ml, 20 ng / ml, 30 ng / ml, 40 ng / ml, 50 ng / ml, respectively, and 50 ng / ml of chondrocytes as the positive control group and BMSCs as the negative control group. The 0ng/ml group was treated with mixed culture without BMP-2 cytokines. Growth factor was added to culture for 24 h, 48 h, 72 h later, the relative cell number (OD) was measured by (MTT) colorimetry, and then the average OD value of BMP-2 at 490nm at different concentrations was calculated and converted to cell relative number. It is concluded that cytokine BMP-2 and chondrocytes have synergistic effects on differentiation and proliferation of bone marrow stromal cells into chondrocytes. Then immunocytochemical staining was used to determine that type 鈪
本文编号：2438233
[Abstract]:Objective to investigate the synergistic effect of bone reactive morphogenetic protein (BMP-2) and chondrocytes on the differentiation of bone marrow stromal stem cells (BMSCs) into chondrocytes. The optimal concentration of BMP-2 on chondrocytes induced by co-culture of BMSCs and chondrocytes was determined. Methods the cultured rabbit BMSCs and chondrocytes were mixed in the same proportion (7:3) in vitro, and cultured in 5. 0 脳 107/ml medium. According to the concentration of different cytokine BMP-2, they were divided into the following groups: 5 ng / ml, 10 ng / ml, 20 ng / ml, 30 ng / ml, 40 ng / ml, 50 ng / ml, respectively, and 50 ng / ml of chondrocytes as the positive control group and BMSCs as the negative control group. The 0ng/ml group was treated with mixed culture without BMP-2 cytokines. Growth factor was added to culture for 24 h, 48 h, 72 h later, the relative cell number (OD) was measured by (MTT) colorimetry, and then the average OD value of BMP-2 at 490nm at different concentrations was calculated and converted to cell relative number. It is concluded that cytokine BMP-2 and chondrocytes have synergistic effects on differentiation and proliferation of bone marrow stromal cells into chondrocytes. Then immunocytochemical staining was used to determine that type 鈪
本文编号：2438233
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