Ikuko Kimura, Motonori Okabe, Masaru Ogasawara and Masayasu Kimura
Department of Chemical Pharmacology, Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama 930-01, Japan
Abstract: With flow cytometry, we investigated the effect of basic fibroblast growth factor (bFGF)- induced competence in subcultured endothelial cells (EC) (4 - 9-passage) of rat thoracic aorta. The cell population in each phase of the cell cycle was determined by a double staining technique with fluorescein isothiocyanate-conjugated mouse monoclonal antibody against the proliferation-associated nucleus antigen Ki-67 and propidium iodide for total DNA content. EC were cultured in medium containing 5% fetal bovine serum (FBS) for 6 days. After serum-starvation for 2 days, the treatment with bFGF (3 - lO ng/ml) for 12 hr promoted the entry of cells into the G1 phase from the G0 phase concentration-dependently. bFGF (10 ng/ml) increased the cell population in the G1 phase by 5% of the total EC, compared with the control culture without bFGF. A further 12 - 15-hr culture with 1% FBS after bFGF treatment promoted the entry of the cell into the S phase. Thus flow cytometric analysis demonstrates that bFGF stimulates the entry of EC into the G1 phase from the G0 phase.
Keywords:
Basic fibroblast growth factor, Flow cytometry, Rat aortic endothelial cell, Ki-67 antigen, Propidium iodide