Flow cytometric cell cycle analysis of cultured brown bear fibroblast cells

Abstract

The aim of this study was to assess by flow cytometry the cell cycle of brown bear fibroblast cells cultured under different growth conditions. Skin biopsies were taken in Cantabria (Spain) from a live, anaesthetized brown bear. DNA analysis was performed by flow cytometry following cell DNA staining with propidium iodide. Serum starvation increased (P < 0.01) the percentage of G0/G1 phase cells (92.7 0.86) as compared to cycling cells (39.7 0.86) or cells cultured to confluency (87.3 0.86). DMSO included for 48 h in the culture significantly increased (P < 0.01) the percentage of G0/G1 phase of the cell cycle at all concentrations used and decreased percentages of S phase in a dose-dependent fashion. Roscovitine increased the G0/G1 phase of the cell cycle (P < 0.01) at 15 mM concentration. Interestingly, the G2/M stage significantly increased at 30 and 50 mM compared to the control and 15 mM (P < 0.02). The cell cycle of brown bear adult fibroblast cells can be successfully synchronized under a variety of culture conditions.