生长因子诱导骨髓间充质干细胞成软骨分化的研究

目的　建立一种体外分离兔骨髓间充质干细胞 (MSCs)的方法 ,诱导其表达软骨细胞表型。方法　抽取兔骨髓 ,经密度梯度离心和粘附分离得到MSCs。用生长因子诱导 ,观察细胞的增殖情况和软骨特异性基质的表达水平。结果　①纤维粘连蛋白促进MSCs贴壁 ,可提高细胞分离率 ;②TGF β1 ,IGF Ⅰ和维生素C结合可促进MSCs增殖 ,表达软骨特异性的Ⅱ型前胶原mRNA和基质蛋白多糖成分。结论　①密度梯度离心结合粘附分离可提高MSCs的分离效率。②TGF β1 ,IGF Ⅰ和维生素C结合诱导可促进MSCs增殖 ,表达软骨细胞表型

Inducing rabbit bone marrow mesenchymal stem cells to express chondrogenic potential by growth factors

Objective To establish a method of isolating rabbit bone marrow mesenchymal stem cells (MSCs) and inducing them to express chondrocyte phenotype. Methods Bone marrow was extracted from the iliac of New Zealand rabbit. MSCs were separated by density centrifugation and fibronectin adhesion. TGF β1, IGF Ⅰ and vitamin C were applied into culture medium to induce proliferation and transformation of MSCs. Procollagen α1(Ⅱ) mRNA and hexuronic acid in culture were detected.Results 1. Adhesion of MSCs to culture flask was enhanced by fibronectin. 2. Cell proliferation and expression of articular cartilage matrix procollagen α1(Ⅱ) mRNA and hexuronic acid were promoted by inducing factors TGF β1,IGF Ⅰ and vitamin C. Conclusion 1.Separation rate of MSCs is enhanced by the combination of density centrifugation and fibronectin adhesion. 2. Cell proliferation and expression of chondrocytic phenotype can be induced by the combination of TGF β1, IGF Ⅰ and vitamin C.