Transfection of bone marrow stromal cells with PcDNA3-hBMP2 and expression detection of hBMP-2 gene

Objective: To explore the feasibility of bone marrow stromal cells (BMSCs) being used as accepted cells of hBMP-2 gene by studying the instantaneous mRNA and its protein expression of hBMP-2 in rabbit's BMSCs, and to make further study on the effects of transfection on biological behavior of these cells. Methods: With the help of the lipofectamine , the cultured BMSCs 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 in situ hybridization and ELISA. Results: After transfection with PcDNA3-hBMP2, hBMP-2 gene's transcribed product- mRNA can be found in the positive cells ,and hBMP-2 proteins can be examined in the culture . The biological behavior of the positive cells unchanged after the target gene was transfected into the cells. Conclusions: The BMSCs could be used as the accepted cells of hBMP-2 gene for expressing and secreting the hBMP-2 into the culture, which can be served as seed cells in bone tissue engineering .