Jin Dan, Zeng Wei-Sen, Pei Guo-Xian, Qi Feng-Ju, Wei Kuan-Hai, Wang Ke, Chen Bin
Department of Orthopedics and Traumatology, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2002 Mar;22(3):222-5.
To detect the expression of human bone morphogenetic protein 7 (hBMP-7) mRNA in rabbit bone marrow-derived mesenchymal stem cells with hBMP-7 gene transfection mediated by retroviral vector.
Retroviral vector for hBMP-7 gene was constructed that was transferred into the packaging cell PT67 mediated by liposome. hBMP-7 gene-positive cell clones were selected with G418 (600 ng/ml) and amplified to obtain the retroviral supernatant containing the target genes that were subsequently used to transfect rabbit bone marrow-derived mesenchymal stem cells (MSCs). HBMP-7 mRNA expression in the MSCs was examined by way of in situ hybridization and reverse transcriptase-polymerase chain reaction.
hBMP-7 retroviral vector was successfully constructed and transferred into PT67 cells with resistance to G418, and after transfection with the recombinant retrovirus, transcription and expression of hBMP-7 mRNA were detected in MSCs.
Rabbit bone marrow stem cells transfected with hBMP gene via retroviral vector can secrete the correspondent protein, indicating the possibility of enhancing the osteogenic capacity of MSCs in the study of bone tissue engineering.
检测逆转录病毒载体介导的人骨形态发生蛋白7（hBMP-7）基因转染兔骨髓间充质干细胞后hBMP-7 mRNA的表达。
构建hBMP-7基因逆转录病毒载体，经脂质体介导转入包装细胞PT67。用G418（600 ng/ml）筛选hBMP-7基因阳性细胞克隆并扩增，获得含目的基因的逆转录病毒上清液，用于转染兔骨髓间充质干细胞（MSCs）。采用原位杂交和逆转录聚合酶链反应检测MSCs中hBMP-7 mRNA的表达。
成功构建hBMP-7逆转录病毒载体并转入对G418有抗性的PT67细胞，用重组逆转录病毒转染后，在MSCs中检测到hBMP-7 mRNA的转录和表达。
经逆转录病毒载体转染hBMP基因的兔骨髓干细胞能分泌相应蛋白，提示在骨组织工程研究中增强MSCs成骨能力的可能性。
