[The biological changes of NIH3T3 cells co-cultured with human bone morphogenetic protein-2 gene transfecting cells].
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.
Wang
Juan
J
Department of Periodontology, College of Stomatology, Nanjing Medical University, Nanjing 210029, China.
Sun
Wei-Bin
WB
Lu
Chun
C
Tang
Gui-Xia
GX
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D018920:Coculture Techniques; D005347:Fibroblasts / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D010012:Osteogenesis; D014162:Transfection
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D000469:Alkaline Phosphatase