【摘要】 目的将间充质干细胞诱导分化为软骨细胞,观察分化后的细胞在单层培养和几丁质支架上培养的差别。方法抽取兔股骨骨髓,密度梯度离心分离BMSCs,用TGF-β1诱导第3代的BMSCs向软骨方向分化,2周后用免疫组化检测Ⅱ型胶原表达情况。将分化后的软骨样细胞传代,用无TGF-β1的培养基分别进行单层培养和接种在几丁质支架上培养2周,再用免疫组化检测Ⅱ型胶原表达情况。结果BMSCs经TGF-β1诱导后,具有软骨细胞特点,然而进行无TGF-β1的继续培养后,单层培养的类软骨细胞很快呈去分化表型,而接种在支架上的细胞仍能较好表达Ⅱ型胶原。结论几丁质支架具有延缓软骨样细胞去分化和老化的作用。更多还原

【Abstract】 Objective To study the difference of the cartilage cells (which were differentiated from mesenchymal stem cells) cultured by means of monolayer culture and chitosan scaffolds respectively.Methods Bone marrow was obtained from the rabbit femora and BMSCs were separated from rabbit bone marrow by density gradient centrifugation. TGF-β1 was used to induce chondrogenic differentiation of the third generation of BMSCs. Two weeks later, Collagen II of the BMSCs was detected by immunohistochemistry, and then the cells induced by TGF-β1 were respectively cultured by means of monolayer culture and chitosan scaffolds for two weeks. The expression of Collagen II was detected again by immunohistochemistry.Results After BMSCs were cultured with TGF-β1, Collagen II detected by immunohistochemistry proved that these cells showed the traits of cartilage cells. But when these cells continued to be cultured without TGF-β1 by means of monolayer culture, they soon showed dedifferentiation of cartilage cells, whereas the cells in the chitosan scaffolds expressed Collagen II normally.Conclusion After BMSCs were induced into cartilage cells, chitosan scaffolds play an important role in delaying aging and dedifferentiation of cartilage cells.更多还原