plRES-骨形态发生蛋白2质粒转染大鼠骨髓间充质干细胞后的持续表达

背景：重组人骨形态发生蛋白2已被广泛应用于骨组织工程治疗骨缺损或骨不连，但是直接应用外源性骨形态发生蛋白2修复骨缺损效果不理想。目的：观察转染plRES-骨形态发生蛋白2的大鼠骨髓问充质干细胞持续合成并分泌骨形态发生蛋白2的情况。方法：全骨髓贴壁法分离培养大鼠骨髓间充质干细胞，脂质体介导下将真核表达质粒plRES-骨形态发生蛋白2导入大鼠骨髓间充质干细胞。结果与结论：ELISA结果显示，随着plRES-骨形态发生蛋白2转染时间的延长，大鼠骨髓问充质干细胞分泌的骨形态发生蛋白2逐渐增多，至转染后10-12d达高峰，转染后15d，仍可见较多骨形态发生蛋白2的表达。说明转染plRES-骨形态发生蛋白2的大鼠骨髓间充质干细胞可持续稳定分泌骨形态发生蛋白2。

Persistent expression of plRES-bone morphogenetic protein 2 plasmid after transfection into rat bone marrow mesenchymal stem cells

BACKGROUND： Recombinant human bone morphogenetic protein has been widely used in bone tissue engineering for treatment of bone defects or bon nonunion. But direct application of exogenous bone morphogenetic protein for repair of bone defects yields unsatisfactory effects. OBJECTIVE： To investigate persistent secretion and expression of bone morphogenetic protein in rat bone marrow mesenchymal stem cells transfected with plRES-bone morphogenetic protein. METHODS： Rat bone marrow mesenchymal stem cells were isolated and cultured by the whole bone marrow adherence method. Under the liposome induction, eukaryotic expression plasmid plRES-bone morphogenic protein 2 was introduced into rat bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION： ELISA results showed that with the prolonged transfection time of plRES-bone morphogenetic protein, bone morphogenetic protein 2 secreted by rat bone marrow mesenchymal stem cells was gradually increased, peaked at 10-12 days after transfection, and was relatively much at 15 days after transfection. These findings suggest that after transfected by plRES-bone morphogenetic protein 2, rat bone marrow mesenchyma stem cells can persistently and stably secrete bone morphogenetic protein 2.