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[The biological changes of NIH3T3 cells co-cultured with human bone morphogenetic protein-2 gene transfecting cells]
Juan Wang1, Wei-Bin Sun, Chun Lu
1Department of Periodontology, College of Stomatology, Nanjing Medical University, Nanjing 210029, China.
Objective:
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.
Methods:
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.
Results:
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.
Conclusions:
Secretive BMP-2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast cells.
