The Expression and Transfection of hBMP-2 Gene in Bone Marrow Stramal Cells

Objective To explore the feasibility of bone marrow stromal cells (MSC) being used as accepted cells of human bone morphogenetic protein-2 (hBMP-2) gene by studying the instantaneous mRNA and protein expression of hBMP-2 in rabbit's MSC, and to make further study on the effects of transfection on biological behavior of these cells. Methods With the help of lipofectamine, the cultured MSCs were transfected with PcDNA3-hBMP2 and the positive cells were screened out by G418. The evidence of successful transfection in these cells could be obtained by PCR,in situ hybridization and ELISA. Results After transfection with PcDNA3-hBMP2, hBMP-2 gene expressed as a 360 bp in MSCs, and its mRNA was found in 15～20% positive cells of MSCs, and the hBMP-2 proteins secreted by the cultured cells were 37mg/3×10 5 cells 1～4 days after transfection, 54ng/3×10 5 cells 11～14 days and 25～28 days after transfection respectively.Conclusion MSC could be used as the receptive cells of hBMP-2 gene and served as seed cells in bone tissue engineering.