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Background Bone morphogenetic proteins (BMPs), which be long to the transforming growth factor beta superfamily, are powerful regulators of cartilage and bone formation. This study investigated the biological change s of NIH3T3 cells incubated with secretive BMP2 that was induced by gene transfe ction through transwell. Methods Eukaryonic expression vector (pcDNA3.1-B2) was transf ered into NIH3T3 cells with Sofast TM,a positive compound transfection age nt. The positive cell clones were selected with G418. The cytoplasmic and extr acellular expressions of BMP2 were determined by immunohistochemical stain and e nzyme-linked immunosorbent assay. NIH3T3 cells were co-cultured with h BMP2 gene transfecting cells through transwell, and the ultrastruct ure, alkaline phosphatase activity and the expression of osteocalcin (the marker of osteogenetic differentiation) changes were observed. Results There were cytoplasmic and extracellular expressions of BMP2 in transfecting NIH3T3 cells. The ultrastructural changes, the high activ ity of alkaline phosphatase and the positive stain of osteocalcin suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfec ting NIH3T3 cells. Conclusion Secretive BMP2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast cells.
Background Bone morphogenetic proteins (BMPs), which be long to the transforming growth factor beta superfamily, are powerful regulators of cartilage and bone formation. This study investigated the biological change of NIH3T3 cells incubated with secretive BMP2 that was induced by gene transfected through Transwell. Methods Eukaryonic expression vector (pcDNA3.1-B2) was transf ered into NIH3T3 cells with Sofast ™, a positive compound transfection age nt. The positive cell clones were selected with G418. The cytoplasmic and extr acellular expressions of BMP2 were determined by immunohistochemical stain and e nzyme-linked immunosorbent assay. NIH3T3 cells were co-cultured with h BMP2 gene transfecting cells through transwell, and the ultrastruct ure, alkaline phosphatase activity and the expression of osteocalcin (the marker of osteogenetic differentiation) changes were observed. Results There were cytoplasmic and extracellular expressions of BMP2 in transfecting NIH3T3 cells. The ul trastructural changes, the high activ ity of alkaline phosphatase and the positive stain of osteocalcin suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfec ting NIH3T3 cells. Conclusion Secretive BMP2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast cells .