骨形成蛋白-7基因逆转录病毒载体的构建及其表达  

Construction and expression of retroviral vector containing bone morphogenetic protein-7 gene

作　　者：祝联[1] 刘伟[1] 陈兵[1] 刘德莉[1] 刘方军[1] 吴娟娟[1] 陆俊弘[1] 曹谊林[1] 

机构地区：[1]上海第二医科大学第九人民医院整复外科上海市组织工程重点实验室,上海200011

出　　处：《细胞与分子免疫学杂志》2003年第4期329-331,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家重点基础研究发展规划 (973)资助 (No.G1 9990 5430 4 )

摘　　要：目的 :实现骨形成蛋白 7(BMP 7)基因在骨髓基质干细胞中的稳定、长久表达。方法 :构建重组BMP 7逆转录病毒表达载体。通过PT6 7包装细胞克隆 ,嘌呤霉素筛选、扩增 ,获得含BMP 7基因的逆转录病毒液 ,直接感染骨髓基质干细胞 ,用免疫组化方法检测BMP 7的表达。结果 :成功地构建了重组BMP 7逆转录病毒表达载体 ,并可在骨髓基质干细胞中表达BMP 7。结论 :重组逆转录病毒表达载体转染的靶细胞可表达BMP 7,为组织工程中骨和软骨种子细胞的研究奠定了基础。AIM: To achieve stable and long term expression of BMP 7 gene in bone marrow stem cells (BMSCs). METHODS: Retrovirus expression vector containing BMP 7 gene was constructed and transfected into packaging cells PT67. After puromycin selection and cells cloning, cells clones producing high level of recombinant virus were obtained. The viruses were used to infect directly BMSCs and the expression of BMP 7 gene in BMSCs was analyzed by immunohistochemical staining. RESULTS: Retrovirus vector containing BMP 7 gene was successfully reconstructed and BMP 7 was expressed in the BMSCs.The transfection rates was about 30%～40%. CONCLUSION: The construction of recombinant retrovirus vector containing BMP 7 gene can provide a reliable tool for the formation of bone or cartilage seed cells in the research of tissue engineering.

关 键 词：重组逆转录病毒载体 骨形成蛋白—7 组织工程 骨髓基质干细胞 

分 类 号：Q782[生物学—分子生物学]