定向诱导人骨髓间质干细胞向软骨细胞分化的初步研究

Study on the directed inducing process of cartilage cells differentiated from human marrow mesenchymal stem cells

目的 采用组织工程方法 ,将人骨髓间质干细胞 (mesenchymalstemcells ,MSC)在体外培养后 ,诱导为软骨细胞 ,为软骨的修复和重建提供条件。方法 根据 5例的预实验经验 ,采用 12例开胸手术所取正常肋骨分别实验。将肋骨剪开 ,用含双抗的不完全培养基反复冲洗骨髓腔得到骨髓液 ,经密度梯度离心获得单个核细胞 ,体外培养、分离获得人骨髓MSC ,经无血清培养诱导液进行诱导培养 ,透射电镜观察细胞外基质形成及超微结构 ,免疫组化检测MSCⅡ型胶原表达。结果 12例的肋骨骨髓MSC分别体外培养 2周后见细胞融合 ,传代培养并进行诱导 ,部分细胞开始由梭形转变为圆形 ,并分泌基质 ,甲苯胺蓝染色呈阳性 ;MSCⅡ型胶原表达阳性。结论 人MSC在体外能诱导为软骨细胞 ,可用于组织工程软骨组织的构建 ,是一种有重要意义的“种子细胞”。

Objective To study the feasibility of constructing tissue engineered cartilage cells by differentiated human bone marrow mesenchymal stem cells(MSC)cultured in vitro Methods The MSC were isolated from the rib by density gradient centrifuge, then induced into chondrogenic differentiation by adding dexamethasone, transforming growth factor β 1(TGF β 1),ascorbic acid, insulin and bovine serum albumin Results Some cells were degenerated and disappeared after cultured in vitro But the residual cells were surviving and have secreted metachromatic matrix ,which is positive with toluidine blue staining Collagen II antigens expression is positive , which is one of the cartilage cells characteristics Conclusion MSC can be induced to cartilage cells in vitro, MSC can be used as functional cells for constructing tissue engineered cartilage