慢病毒介导胰岛素生长因子1促进人骨髓间充质干细胞的成骨分化  

Effects of osteogenetic differentiation of human insulin-like growth factor-1 on human bone marrow stem cells transfected

作　　者：冯尚祥 吕成昱[2] 隋爱华[3] 申成凯 张海宁[2] 

机构地区：[1]青岛大学医学院附属医院,山东省青岛市266003 [2]青岛大学医学院附属医院关节外科,山东省青岛市266003 [3]青岛大学中心实验室,山东省青岛市266003 [4]青岛市胶州中心医院骨科,山东省青岛市266003

出　　处：《中国组织工程研究》2015年第B05期39-42,共4页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目（81171774.81272056）

摘　　要：背景：目前慢病毒介导胰岛素生长因子1基因作用于人骨髓间充质干细胞的研究还处在初级阶段。目的：构建携带胰岛素生长因子1基因的慢病毒载体，观察其在人骨髓间充质干细胞中的表达以及其对成骨分化的影响。方法：应用重组慢病毒技术将胰岛素生长因子1基因和绿色荧光蛋白转染至体外培养第3代人骨髓间充质干细胞（实验组），以空白的慢病毒转染的人骨髓间充质干细胞和未做处理的人骨髓间充质干细胞分别作为对照组和空白组进行对比。结果与结论：慢病毒转染3d后，对照组和实验组的转染效率分别可达95％和90％以上。Westernblot检测结果和激光共聚焦显微镜下观察显示，胰岛素生长因子1在人骨髓间充质干细胞中成功表达。说明胰岛素生长因子1基因转染对人骨髓间充质干细胞成骨能力具有促进作用。BACKGROUND： Research on bidirectional regulation effects of human insulin-like growth factor-1 （hlGF-1） gene induction on stem cells is still in primary phase currently. OBJECTIVE： Lentivirus vector carrying hlGF-1 gene were used to transfect human bone marrow mesenchymal stem cells, and then to observe the expression of target gene and differentiation of those cells. METHODS： The recombinant lentiviral vectors carrying hlGF-1 gene and green fluorescent protein were constructed with recombinant lentiviral technology, and then the recombinant lentiviral vectors were used to transfect the passage 3 human bone marrow mesenchymal stem cells in vitro cultured （transfection group）. The bone marrow mesenchymal stem cells transfected with lentivirals were as control （control group）. RESULTS AND CONCLUSION： At 3 days after trasfection, the green fluorescence bone marrow mesenchymal stem cells could be observed under fluorescence microscope, and the transfection efficiency of control group was over 95%,while the transfection group was over 90%. Confocal laser scanning microscope and Western blot results showed that hlGF-1 gene have the positive effect of promoting expression in cellular and protein level of human bone marrow mesenchymal stem cells.

关 键 词：骨髓间充质干细胞 胰岛素生长因子1 基因转染 成骨分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]