BMP7腺病毒基因转染对骨髓基质干细胞生物学的影响

目的 探讨BMP7腺病毒基因转染对骨髓基质干细胞(BMSCs)的体外、体内的生物学功能影响.方法 抽取羊的骨髓,分离、培养骨髓基质干细胞.用重组骨形成蛋白7腺病毒(adenovirus bone morphogenetic protein 7;Adeno-BMP7)转染(M.O.I=100)70%～80%融合时的第二代细胞,三天后分别进行透射电镜观察、流式细胞仪检测、Western-blot、钙结节染色以及珊瑚和细胞复合物皮下回植.结果 在体外:细胞超微结构观察显示Adeno-BMP7基因转染后的BMSCs的物质合成代谢功能活跃;流式细胞仪检测表明Adeno-BMP7基因转染对BMSCs的细胞周期无明显影响;Western-blot检测证明转染Adeno-BMP7后的BMSCs有BMP7在蛋白水平上的表达;染色表明转染Adeno-BMP7后的BMSCs可形成较大的钙结节.在体内:转染Adeno-BMP7的BMSCs可明显促进新骨的形成,与没转染Adeno-BMP7的BMSCs有差异.结论 Adeno-BMP7转染可促进BMSCs的体外成骨定向分化,Adeno-BMP7转染后的BMSCs具有更强的体内成骨能力.

Effect of Adeno-BMP7 Transfection on the Biology of BMSCs In Vitro and In Vivo

Objective To study the effect of Adeno-BMP7 transfection on the biology of Bone marrow stromal cells (BMSCs) In vitro and In vivo. Methods Bone marrow was obtained from goat, and BMSCs were isolated and cultured at the second passage. Once cells have attached and formed a monolayer with 70-80% confluency, Adeno-BMP7 (M.O.I.=100) was added to the cells. Three days later, the following procedures were performed to study: cellular ultra-structure observated by transmission electromi croscope, cell cycle monitored by fluorescent-activated cell sorter (FACS), calcium node by staining, replant subcutaneously with cell-coral compound. Result In vitro: active substance metabolism was found in the Adeno-BMP7 infected BMSCs by transmission electromicroscope, FACS analysis found no affect on the cell cycle, and bigger calcium nodes were observed by staining. In vivo, new bone formiation was stimulated with Adeno-BMP7 transfected when evaluated by X-ray and histology with difference from the control BMSCs. Conclusion Adenovirus-BMP7 infection can induce BMSCs to osteogenic in vitro, and the infected cells show much stronger osteogenic ability in vivo.