تاثير كاهش سرم و تراكم كامل سلولي بر هم زمان سازي سيكل سلولي و وقوع آپوپتوز در فيبروبلاست هاي درمي بز

Effect of Serum Starvation and Full Confluence on Cell Cycle Synchronization and Apoptosis of Goat Dermal Fibroblast

Objective: We examine the effects of serum starvation, culture 10 confluence, and full confluence on cell cycle characteristics and apoptosis of goat dermal fibroblast cells. Materials and Methods: Fibroblast cells were obtained from the ear of a female goat , 1.5 years of age. The following experimental groups were analyzed for fibroblast cells : 1. cells confluent for 72h, 2. cells starved for 48 and 72h. Results: Analysis of cell cycle distribution by flow cytometry showed that 4.56, 51.88 of normal cycling cells were at the GO, G1 phases, respectively. Serum starvation for 48 and 72h arrested cells at the GO/G1 phase (p<0.05). 91.53% of full confluent cells were at GOI G1 stage, but in contrast to the serum starved group, this high percentage of GO/G1 cells was mainly associated with G1 cells. When DNA synthesis was detected by BrdU incorporation 19.80% of normally growing cells were in S phase. The percentage of cells in S phase changed significantly among 48 and 72h serum strarvation and full confluent groups. Under normal culture conditions, 6.67% of cells underwent apoptosis . Serum starvation for 48 and 72 hours caused early apoptosis in 8.91 and 39.83 of cells , respectively. Treatment with full confluen ce 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. Although serum starvation for 72 hours effectively arrested cells at the GO/G1 stages, it significantly increased cell apoptos is.