中文摘要：

目的：探讨人软骨细胞培养上清诱导冻存人骨髓充质干细胞向软骨细胞分化的可行性。方法：取进行全髋关节置换术老年患者的骨髓和软骨组织,利用密度梯度离心法、全骨髓培养法分别培养骨髓间充质干细胞,冻存备用。培养软骨细胞,观察细胞生长,收集软骨细胞培养上清。复苏冻存的人骨髓间充质干细胞,观察复苏后细胞生长状态。利用收集的软骨细胞培养上清对复苏间充质干细胞进行定向诱导,诱导培养2周,观察细胞外观表型变化,II型胶原免疫组化检测诱导后人骨髓间充质干细胞II型胶原的表达。结果：密度梯度离心法与全骨髓培养法均可分离获得人骨髓间充质干细胞,原代生长前者优于后者。复苏细胞仍进行可传代,与正常生长骨髓间充质细胞无明显差异,均可传至第8代。软骨细胞培养上清诱导2周后,细胞形状向圆形,多角形转变,冻存骨髓间充质干细胞II型胶原免疫组化检测II型胶原表达阳性。结论：老年人骨髓间充质干细胞仍具有向软骨细胞转化的能力,冻存不影响其转化能力。

英文摘要：

Objective： To explore the feasibility of human cryopreserved bone marrow mesenchymal stem cells（BMSCs） induced by the supernatants of chondrocytes to chondrocytes.Methods： Got the marrow and cartilage tissue from old people who underwent total hip replacement.BMSCs were separated respectively with density gradient centrifugation and whole bone marrow culture method.Human hyaline cartilage was dissected,digested,and chondrocytes were cultured in vitro.View the chondrocytes under microscope and collected the supematants of chondrocytes.The cryopreserved BMSCs were cultured with supematants from chondrocytes.Two weeks later,identify the changes of phenotype of the cells,and immunohistochemistry for type II collagen were used to detect the possible changes of BMSCs.Results： BMSCs could be harvested using both of these two methods,but the primary cells got from density gradient centrifugation were better.There was no difference in generation between the normal and cryopreserved BMSCs,both could go down to 8th generation.Two weeks later,extrinsic feature of cryopreserved BMSCs turned to polygon and circular.Immunohistochemistry staining showed some cells of BMSCs expressed type II collagen.Conclusion： Cryopreserved BMSCs got from old people could be induced by supematants from chondrocytes to differentiate to chondrocytes.