BMP-7基因修饰的骨髓间充质干细胞在小鼠肾脏内的表达

Expression of BMP-7 gene modified bone marrow mesenchymal stem cells in the kidneys of mice

目的构建BMP-7基因腺相关病毒,转染骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs),移植入小鼠体内,观察其在肾脏的定植情况。方法采用分子克隆技术,构建重组质粒pAAV-BMP-7。采用磷酸钙沉淀法,重组质粒与pAAV-RC、pHelper共转染HEK-293细胞,产生具有传染性的病毒颗粒。将此病毒颗粒转染BMSCs后,经尾静脉注射入小鼠体内,免疫组化方法检测其在肾脏的定植情况。结果成功构建了BMP-7基因腺相关病毒,可以高效转染BMSCs,移植入小鼠体内后,该细胞可在肾脏内定植。结论BMSCs能稳定、高效表达外源基因BMP-7,并可在肾脏内定植。

Objective To investigate the expression of BMP-7 gene modified bone marrow mesenchymal stem cells（BMSCs） after mice were exposed to ^60Co gamma ray. Methods The recombinant adeno-associated virus vectors encoding BMP-7 （pAAV-BMP-7） were constructed with the technique of molecular cloning. The recombinant expression plasmid were co-transfected into HEK-293 cells with pHelper and pAAV-RC by calcium-phosphate precipitation method. Recombinant AAV viral particles from infected HEK-293 cells were used to infect BMSCs. The infected MMSCs were injected into mice through caudal vein after the BALB/c mice were exposed to 8.0 Gy ^60Co γ/ ray. BMP-7 expressions were observed by immunocytochemical staining in the kid- neys of control mice only exposed to γ/ray and mice transfected with BMSCs or BMP-7 gene modified BMSCs. Results The recombinant AAV vectors of BMP-7 gene were constructed and expressed in BMSCs successfully. The infected BMSCs could lodge in the kidneys. Conclusion These genetically modified BMSCs might be used for further research on gene therapy of renal diseases.