Document Type : Original Article
Embryology Department, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Genetic Department, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Stem Cells and Developmental Biology Department, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Science, Yazd, Iran
Objective: We examine the effects of serum starvation, culture to 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 G0, G1 phases, respectively. Serum starvation for 48 and 72h 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 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 confluence 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 G0/G1 stages, it significantly increased cell apoptosis.