【摘要】 目的构建携带骨形成蛋白-2(BMP-2)与碱性成纤维细胞生长因子(bFGF)双基因过表达的慢病毒载体,观察其对兔骨髓间充质干细胞(BMSCs)增殖的影响。方法体外培养兔BMSCs,构建携带b FGF和BMP-2双基因的重组慢病毒载体,转染至BMSCs诱导两种基因过表达;流式细胞仪检测转染效率;Western blot方法验证双基因转染模型构建成功;MTT方法检测细胞增殖水平。结果慢病毒以最佳感染复数(MOI)100转染BMSCs时转染效率最佳,Western blot检测到转染后细胞bFGF和BMP-2蛋白表达量明显比对照增加(P<0.05);MTT结果发现,过表达bFGF和BMP-2的BMSCs细胞的增殖水平增加(P<0.05)。结论 bFGF和BMP-2双基因共表达重组腺病毒载体促进BMSCs细胞增殖。更多还原

【Abstract】 Objective To construct bone morphogenetic protein 2(BMP-2) and basic fibroblast growth factor(bFGF) double gene lentivirus vectors, and to observe its effect on the proliferation of bone marrow mesenchymal stem cells(BMSCs) in rabbit. Methods Rabbit BMSCs were cultured in vitro. The recombinant lentivirus vectors carrying b FGF and BMP-2 were constructed and transfected into BMSCs to induce overexpression of two genes. The transfection efficiency was detected by flow cytometry. Western blot was used to verify the success of the two-gene transfection model. The cell proliferation was detected by MTT. Results Lentivirus was transfected with the optimal complex number(MOI) 100 for BMSCs. The expression level of bFGF and BMP-2 proteins was significantly increased in transfected cells than in the control group(P<0.05). MTT results showed that the proliferation rates of BMSCs with overexpression of bFGF and BMP-2 increased(P<0.05). Conclusion The combined b FGF and BMP-2 recombinant adenovirus vector promoted the proliferation of BMSCs cells.更多还原