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骨髓间充质干细胞(BMSCs)可分化成多种细胞。已有研究表明,局部微环境影响细胞的生长、分化等功能,不同的细胞共培养可为细胞提供特定的环境。因此,本实验将大鼠BMSCs与大鼠韧带成纤维细胞间接共培养不同时间段后,检测其I型、III型胶原和韧粘素-C的mRNA表达以及I型、III型胶原的蛋白合成量。结果显示,体外培养6d,间接共培养组BMSCsI型胶原mRNA的表达量(3.9±0.2)是对照组(1.9±0.3)的2.0倍;间接共培养组BMSCsIII型胶原的表达量(1.9±0.2)是对照组(0.8±0.1)的2.2倍,而韧粘素-C的mRNA表达量两组间无明显差异。体外培养12d,间接共培养组BMSCs的I型和III型胶原mRNA表达量继续升高,它们的蛋白含量分别从对照组的12.4±0.8ng/μg和5.0±0.4ng/μg,增加到13.6±1.3ng/μg和5.9±0.5ng/μg;韧粘素-C的mRNA表达量(0.14±0.02)为对照组(0.07±0.02)的2.0倍。结果提示与韧带成纤维细胞间接共培养,可以促进BMSCsI型、III型胶原蛋白和韧粘素-C的合成。
Bone marrow mesenchymal stem cells (BMSCs) differentiate into a variety of cells. Studies have shown that the local microenvironment affects cell growth and differentiation functions, and different cell co-culture can provide cells with a specific environment. Therefore, in this experiment, rat BMSCs were indirectly co-cultured with rat ligament fibroblasts for different time periods, and the mRNA expression of type I, type III collagen and tenascin-C and the protein synthesis of type I and type III collagen the amount. The results showed that the expression of BMSCs collagen type Ⅰ mRNA (3.9 ± 0.2) was 2.0 times that of the control group (1.9 ± 0.3) after cultured for 6 days in vitro. The expression of BMSCs type III collagen in the co-culture group was 1.9 ± 0.2 Which is 2.2 times that of the control group (0.8 ± 0.1), while there is no significant difference between the two groups in the mRNA expression of tenascin-C. After cultured for 12 days, the expression of type I and type III collagen mRNAs in BMSCs of Indirect co-culture group continued to increase, and their protein content increased from 12.4 ± 0.8ng / μg and 5.0 ± 0.4ng / μg in control group to 13.6 ± 1.3 ng / μg and 5.9 ± 0.5 ng / μg, respectively. The mRNA expression level of tenascin-C (0.14 ± 0.02) was 2.0 times that of the control group (0.07 ± 0.02). The results suggest that indirect co-culture with ligament fibroblasts can promote the synthesis of BMSCs type I, type III collagen and tenascin-C.