The biological changes of NIH3T3 cells co-cultured with human bone morphogenetic protein-2 gene transfecting cells
Wang, J.; Sun, W.-B.; Lu, C.; Tang, G.-X.
Zhonghua Kou Qiang Yi Xue Za Zhi 41(2): 77-80
2006
ISSN/ISBN: 1002-0098 PMID: 16640928 Document Number: 599711
To investigate the ultrastructure and the alkaline phosphatase (ALP) activity changes of NIH3T3 cells incubated with secretive human bone morphogenetic protein-2 (hBMP-2) that is induced by gene transfection through transwell system. Eukaryotic expression vector (pcDNA3.1-B2) was transduced into NIH3T3 cells by Sofast, a positive compound transfection agent. The positive cell clones were selected with G418. The cytoplasmic and extracellular expression of BMP-2 in the NIH3T3 cells were determined by immunohistochemical and enzyme-linked immunosorbent assay (ELISA). NIH3T3 cells were co-cultured with hBMP-2 gene transfecting cells through transwell system, and the ultrastructure and ALP activity (the markers of osteogenetic differentiation) changes were observed. There were cytoplasmic and extracellular expression of BMP-2 in transfecting NIH3T3 cells. The ultrastructure changes and the high expression of ALP suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfecting NIH3T3 cells. Secretive BMP-2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast cells.