Cell Journal (Jan 2009)
Effect of Serum Starvation and Full Confluence on Cell Cycle Synchronization and Apoptosis of Goat Dermal Fibroblast
Abstract
Objective: We examine the effects of serum starvation, culture to confluence, and full confluenceon cell cycle characteristics and apoptosis of goat dermal fibroblast cells.Materials and Methods: Fibroblast cells were obtained from the ear of a female goat, 1.5years of age. The following experimental groups were analyzed for fibroblast cells: 1. cellsconfluent for 72h, 2. cells starved for 48 and 72h.Results: Analysis of cell cycle distribution by flow cytometry showed that 4.56, 51.88 ofnormal cycling cells were at the G0, G1 phases, respectively. Serum starvation for 48 and72h arrested cells at the G0/G1 phase (p<0.05). 91.53% of full confluent cells were at G0/G1 stage, but in contrast to the serum starved group, this high percentage of G0/G1 cellswas mainly associated with G1 cells. When DNA synthesis was detected by BrdU incorporation19.80% of normally growing cells were in S phase. The percentage of cells in S phasechanged significantly among 48 and 72h serum strarvation and full confluent groups. Undernormal culture conditions, 6.67% of cells underwent apoptosis. Serum starvation for 48 and72 hours caused early apoptosis in 8.91 and 39.83 of cells, respectively. Treatment with fullconfluence for 72 hours did not increase the number of apoptotic cell significantly (6.39%).Serum starvation for 72 hours increased early apoptosis significantly (p<0.05).Conclusion: Goat dermal fibroblasts at various stages of the cell cycle were effectively synchronized;that could be beneficial for somatic cell nuclear transfer in this species. Althoughserum starvation for 72 hours effectively arrested cells at the G0/G1 stages, it significantlyincreased cell apoptosis.
