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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。
OBJECTIVE: To achieve the stable and long-term expression of bone morphogenic protein 7 (BMP 7) gene in bone marrow stromal cells. Methods: Recombinant BMP 7 retrovirus expression vector was constructed. The cell clones were packaged by PT6 7, puriunin was screened and amplified to obtain BMP7 gene-containing retrovirus. The recombinant plasmid was directly infected into bone marrow stromal cells and the expression of BMP7 was detected by immunohistochemistry. Results: Recombinant BMP 7 retroviral vector was successfully constructed and BMP 7 was expressed in bone marrow stromal cells. CONCLUSION: The target cells transfected with recombinant retroviral vector can express BMP7, which lays the foundation for the research of bone and cartilage seed cells in tissue engineering.