CAJ | 학술논문

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。
배경：목전적음다연구도시통과체외가입생장인자유도골수간충질간세포분화，단해방법세포인자용량대，성본고，수착배양차수적증가，기성연골잠능명현강저。 　　목적：혼합배양인관절액여인골수간충질간세포，관찰공배양계통대인골수간충질간세포정향유도분화적영향。방법：채용전골수、첩벽배양법배양화분리인골수간충질간세포，무균조작하추취건강지원자적슬관절액，장기여P3대세포혼합용우실험，분위3조，관절액+완전배양기，관절액+인골수간충질간세포+완전배양기，인골수간충질간세포+완전배양기。매천재도치현미경하관찰세포형태변화화생장정황，분별우유도후제7，14，21천행갑분알람염색검측화Ⅱ형효원면역조화염색검측。 　　결과여결론：인관절액여인골수간충질간세포혼합배양후，세포증식속도감만，유장사형변위다각형、타원형，세포외기질정갑분알람이염성양성，Ⅱ형효원면역조화염색양성。설명관절액대인골수간충질간세포향연골세포분화유정성촉진작용，관절액중가능함유촉인골수간충질간세포향연골세포방향분화적물질。
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. 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 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.