转基因技术在组织工程体外软骨组织培养中的应用

Application of transgenic technology in tissue engineering to culture cartilage in vitro

目的:研究骨形态发生蛋白-2重组腺病毒(Ad-BMP-2)转基因对骨髓基质细胞(mesenchymalstemcell,MSC)与纤维蛋白凝胶构成的复合物体外培养的影响。方法:Ad-BMP-2转染原代培养的MSC,通过细胞生长曲线、免疫组化、甲苯胺蓝染色透射电镜观察转基因MSC的BMP-2和CollagenⅡ及蛋白多糖表达。结果:转基因MSC经纤维蛋白体外三维培养后免疫组化和甲苯胺蓝染色呈阳性;电镜显示细胞生长良好,有基质合成。结论:Ad-BMP-2能够通过促进MSC分泌BMP-2诱导CollagenⅡ和蛋白多糖的表达,在纤维蛋白三维支架中形成软骨基质。

AIM:To study the effect of transgenic of bone morphogenetic protein 2 adenoviral vector Ad BMP 2 on the cultivation in vitro of compound of fibrin gel and mesenchymal stem cell (MSC). METHODS:MSC was transferred by Ad BMP 2.BMP of the transgeneic MSC,CollagenⅡ and the expression of proteoglycan were observed by the growth curve of cells,Immunohistochemical method and microscope,Aniline dyeing. RESULTS:Toluidine blue staining and immunohistochemical method showed that transgeneic MSCs were positive after in vitro three dimensional culture in fibrin.Electron microscope showed the cells were well grown and there were Stroma synthesized. CONCLUSION:Ad BMP 2 can induce the expression of Collagen Ⅱ and proteoglycan in MSC by secreting BMP 2,and can reconstruct articular cartilage cultured in fibrin gel.