人关节液促进骨髓间充质干细胞的定向分化  

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

作　　者：华强[1] 吴佳奇[2] 钟传山 刘宗超[2] 严光建[1] 熊小天[2] 崔小明[2] 

机构地区：[1]成都体育学院附属体育医院骨科,四川省成都市610041 [2]泸州医学院附属中医院骨科,四川省泸州市646000

出　　处：《中国组织工程研究》2014年第10期1490-1495,共6页Chinese Journal of Tissue Engineering Research

基　　金：四川省教育厅资助项目(07ZB035)~~

摘　　要：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。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 wil 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 fol owing 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. 〈br〉 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 col agen II staining＆amp;nbsp;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.

关 键 词：干细胞 骨髓干细胞 骨髓间充质干细胞 关节液 软骨细胞 细胞培养 软骨组织工程 Ⅱ型胶原 共培养 

分 类 号：R394.2[医药卫生—医学遗传学]