【摘要】 目的:应用转化型生长因子TGF-β1,体外诱导骨髓间质干细胞(MSCs)向成软骨细胞表型分化,探讨其作为组织工程化软骨种子细胞的可行性。方法:由大鼠骨髓中分离出MSCs,体外传代培养,取第3代细胞通过TGF-β1、地塞米松和维生素C诱导向软骨细胞分化。诱导后14d观察细胞形态变化,进行阿辛蓝染色检测软骨基质的分泌、免疫组化检测细胞Ⅱ型胶原的表达,采用Western-blot和RT-PCR检测诱导前后成软骨相关的SOX9、蛋白聚糖与Ⅱ型胶原的表达。结果:诱导后阿辛蓝染色示糖胺聚糖均匀分布于基质中,免疫组化染色示基质中Ⅱ型胶原表达阳性。RT-PCR检测示成软骨相关的SOX9、蛋白聚糖、Ⅱ型胶原mRNA表达阳性。Western-blot印迹检测示细胞诱导后Ⅱ型胶原蛋白表达阳性。结论:MSCs在特定培养液的诱导下可向软骨细胞表型分化,并能分泌软骨细胞特异性基质,可成为软骨组织工程理想的种子细胞来源。更多还原

【Abstract】 Objective To observe the effects of transforming growth factor-beta 1 (TGF-?茁1) on differentiation into chondrogenic phenotype from bone marrow stromal cells(BMSCs) in rats and to explore the feasibility of the seed cells of tissue-engineering chondrogenesis. Methods BMSCs were isolated from rat bone marrow,cultrued and passaged. The cultured BMSCs at the 3rd passage were induced to differentiate into chondrocytes in induction medium containing TGF-?茁1, desamethason and vitamin C. Fourteen days later, the matrix of cartilage cells and cartilage specific collagen Ⅱ was detected by alcian blue staining and immunohistochemistry, respectively. The expression of SOX9, agrrecan and collagen Ⅱ in the induced MSCs cells were also dectected by Western-blot and TR-PCR. Results After the induction of induced medium containing TGF-?茁1, bone marrow stromal cells could differetiate into cartilage cells. Histological staining of proteoglycan with Alcian blue revealed glycosaminoglycan deposition and immunohistrochemical staining revealed the positive expression of cartilage specific collagen Ⅱ in the typical cartilage extracellular matrix. RT-PCR gene expression analysis of characteristic chondrogenic related genes, such as SOX9, agrrecan as well as collagen Ⅱ were positive, and Western-blot dectection confirmed collagen Ⅱ expression in the matrix of cartilage cells after 14 days induction. Conclusion MSCs from the bone marrow of rats can differentiate into chongrogenic phenotype when cultured in a defined medium. MSCs can likely be served as the optimal source of autogenous seed cells for cartilage tissue engineering.更多还原