The biological changes of NIH3T3 cells co-cultured with human bone morphogenetic protein-2 gene transfecting cells.
- Author:
Juan WANG
1
;
Wei-Bin SUN
;
Chun LU
;
Gui-Xia TANG
Author Information
- Publication Type:Journal Article
- MeSH: Alkaline Phosphatase; metabolism; Animals; Bone Morphogenetic Protein 2; genetics; Coculture Techniques; Fibroblasts; cytology; ultrastructure; Humans; Mice; NIH 3T3 Cells; Osteogenesis; Transfection
- From: Chinese Journal of Stomatology 2006;41(2):77-80
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSEukaryotic 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.
RESULTSThere 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.
CONCLUSIONSSecretive BMP-2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast cells.
