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Peptides from Phage Display Library Modulate Gene Expression in Mesenchymal Cells and Potentiate Osteogenesis in Unicortical Bone Defects
Published on: December 10, 2010
[Expression of human bone morphogenetic protein 7 mRNA after the gene transfection in rabbit bone marrow-derived
Dan Jin1, Wei-Sen Zeng, Guo-Xian Pei
1Department of Orthopedics and Traumatology, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Objective:
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.
Methods:
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.
Results:
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.
Conclusion:
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.
