OBJECTIVE: To elucidate the mechanism of bone morphogenetic protein (BMP) on cell differentiation and to provide basis for BMP gene therapy. METHODS: A phagemid expression vector for human BMP2 (pBK-B2) was constructed and transfected into the NIH3T3 cells by using Lipofect AMINE. Positive cell clones were selected with G-418. The stable transfection, expression and secretion of hBMP2 were determined by in situ hybridization, immunohistochemistry and sandwich-in ELISA methods. The proliferativity of the transfected cells were assayed by methabenzthiazuron (MTT) method. Alkaline phosphatase (ALP) and osteocalcin (OC) production were also measured in the transfected cells. RESULTS: The results showed that cell proliferation was inhibited after the transfection with hBMP2 gene. But the transfected cells showed increased ALP activity and OC production. CONCLUSION: These results indicate that BMP2 is expressed stably and efficiently in the NIH3T3 cells and is involved in inducing differentiation of NIH3T3 cells into osteoblast-like cells.
[Effects of human BMP2 gene transfection on NIH3T3 cells in vitro] Publishing Authors By Initials
