人关节液促进骨髓间充质干细胞的定向分化

背景：目前的众多研究都是通过体外加入生长因子诱导骨髓间充质干细胞分化，但该方法细胞因子用量大，成本高，随着培养次数的增加，其成软骨潜能明显降低。 目的：混合培养人关节液与人骨髓间充质干细胞，观察共培养系统对人骨髓间充质干细胞定向诱导分化的影响。 方法：采用全骨髓、贴壁培养法培养和分离人骨髓间充质干细胞，无菌操作下抽取健康志愿者的膝关节液，将其与P3代细胞混合用于实验，分为3组，关节液+完全培养基，关节液+人骨髓间充质干细胞+完全培养基，人骨髓间充质干细胞+完全培养基。每天在倒置显微镜下观察细胞形态变化和生长情况，分别于诱导后第7，14，21天行甲苯胺蓝染色检测和Ⅱ型胶原免疫组化染色检测。 结果与结论：人关节液与人骨髓间充质干细胞混合培养后，细胞增殖速度减慢，由长梭形变为多角形、椭圆形，细胞外基质呈甲苯胺蓝异染性阳性，Ⅱ型胶原免疫组化染色阳性。说明关节液对人骨髓间充质干细胞向软骨细胞分化有正性促进作用，关节液中可能含有促人骨髓间充质干细胞向软骨细胞方向分化的物质。中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程全文链接：

Human synovial fluid promotes directed differentiation of bone marrow mesenchymal stem cells

BACKGROUND:Nowadays, growth factors are commonly used to induce bone marrow mesenchymal stem cells. However, this is a high-cost method with a great amount of growth factors. In addition, the chondrogenic potential of bone marrow mesenchymal stem cells will decrease significantly with increasing times of culture. OBJECTIVE:To observe the directed differentiation of bone marrow mesenchymal stem cells co-cultured with human synovial fluid. METHODS:Human bone marrow mesenchymal stem cells were isolated and cultured by adherence screening method. The synovial fluid of the knee was aspirated from healthy volunteers by aseptic operation. Passage 3 human bone marrow mesenchymal stem cells were co-cultured with the following media: synovial fluid+complete medium; synovial fluid+bone marrow mesenchymal stem cells+complete medium; bone marrow mesenchymal stem cells+complete medium. The morphology and growth of the cells were observed under an inverted microscope every day. At days 7, 14 and 21 of induction, toluidine blue staining and immunocytochemical staining were performed. RESULTS AND CONCLUSION:After co-culture with human synovial fluid, human bone marrow mesenchymal stem cells proliferated slowly, and varied from fusiform to oval or polygonal; toluidine blue and collagen II staining were positive. These findings indicate that the synovial fluid has a positive role in the chondrogenic differentiation of bone marrow mesenchymal stem cells. The synovial fluid may contain substances that promote the chondrogenic differentiation of bone marrow mesenchymal stem cells.