骨形态发生蛋白7转染骨髓间充质干细胞再生骨组织

Bone tissue regeneration of bone morphogenetic protein-7-transfected bone marrow mesenchymal stem cells

背景:骨形态发生蛋白7具有促进骨髓间充质干细胞分裂增殖的作用,在诱导新骨形成方面有重要的研究意义。目的:观察携带人骨形态发生蛋白7慢病毒载体转染后骨髓间充质干细胞骨形态发生蛋白7蛋白及mR NA的表达。方法:提取并培养大鼠骨髓间充质干细胞,通过形态学和细胞表面标记进行鉴定;构建携带人骨形态发生蛋白7的慢病毒载体,转染大鼠骨髓间充质干细胞,提取RNA后采用RT-PCR的方法检测骨形态发生蛋白7mR NA水平相对表达量的变化,Western blotting检测骨形态发生蛋白7蛋白水平相对表达量的变化。结果与结论:成功构建了携带骨形态发生蛋白7基因的慢病毒载体PLV-sfG FP(2A)-BMP7,重组载体质粒成功转染至大鼠骨髓间充质干细胞中,转染组的骨形态发生蛋白7 m RNA和蛋白表达高于空载体组和空白组,差异有显著性意义(P<0.05),说明外源性骨形态发生蛋白7基因被有效整合到骨髓间充质干细胞并进行表达。

BACKGROUND： Bone morphogenetic protein-7 can promote proliferation of bone marrow mesenchymal stem cells and has important significance in terms of inducing new bone formation.OBJECTIVE： To observe the protein and m RNA expression of bone morphogenetic protein-7 in bone marrow mesenchymal stem cells transfected with bone morphogenetic protein-7.METHODS： Rat bone marrow mesenchymal stem cells were isolated and cultured, and then identified by the morphology and cell surface markers. Lentiviral vectors carrying bone morphogenetic protein-7 were constructed to transfect bone marrow mesenchymal stem cells. After total RNA extraction, the protein and mR NA expression of bone morphogenetic protein-7 was detected by RT-PCR and western blot methods.RESULTS AND CONCLUSION： PLV-sf GFP（2A）-BMP7 recombinant plasmids were successfully constructed and transferred into rat bone marrow mesenchymal stem cells. The mR NA and protein expression of bone morphogenetic protein-7 in the transfection group was significantly higher than that in the blank vector and blank groups（P〈0.05）, indicating exogenous bone morphogenetic protein-7 gene is effectively integrated into the bone marrow mesenchymal stem cells.