论文部分内容阅读
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。
Objective: To investigate the feasibility of human chondrocyte culture supernatant induced chondrogenic stem cells differentiated into chondrocytes. Methods: Bone marrow and cartilage tissues of elder patients undergoing total hip arthroplasty were selected. Bone marrow mesenchymal stem cells were cultured by density gradient centrifugation and whole bone marrow culture respectively. Chondrocytes were cultured, cell growth was observed, and chondrocyte culture supernatants were collected. The cryopreserved human bone marrow mesenchymal stem cells were resuscitated and the cell growth status was observed after resuscitation. The collected supernatant of chondrocytes was used to induce the recovery of mesenchymal stem cells, and the cells were induced to grow for 2 weeks to observe the appearance of cells. The expression of type II collagen in human bone marrow mesenchymal stem cells was detected by immunohistochemistry . Results: Human bone marrow mesenchymal stem cells could be isolated by density gradient centrifugation and whole bone marrow culture. The primary growth was better than the latter. Resuscitation cells were still passaged, and normal growth of bone marrow mesenchymal cells no significant difference, can be passed to the 8th generation. The chondrocyte culture supernatant was induced for 2 weeks, the cell shape turned round and polygonal, and the expression of type II collagen was detected by cryopreserved type II collagen of bone marrow mesenchymal stem cells. CONCLUSION: Bone marrow-derived mesenchymal stem cells from old people still have the ability to transform into chondrocytes, and the cryogenic storage does not affect their transformation ability.