Mimosine As Well As Serum Starvation Can Be Used for Cell Cycle Synchronization of Sheep Granulosa Cells

Abstract

This study was evaluated the effect of different synchronization protocols such as serum starvation for 1–3 days, confluency and chemical inhibitors on synchronization accuracy at G0/G1, apoptosis, and DNA synthesis in sheep granulosa cells. The cells were obtained from ovarian antral follicles of slaughtered sheep and used at first and fifth passages. Flow cytometry analysis showed that confluent cells, serum starvation for 24, 48, and 72 hours, and mimosine treatment significantly increased G0/G1 phase cells when compared to normally growing cells ( P < 0.05 ). Nocodazole treatment increased the cell population in the G0/G1 stage when compared with the control group but did not change the G2/M stage population. Treatment of cells with mimosine, nocodazole, and serum starvation in three groups resulted in proliferation arrest ( P < 0.05 ). Serum starvation for 72 hours significantly promoted apoptosis in granulosa cells ( P < 0.05 ). The results of the primary culture and 5th passage were the same. The use of 48-hour serum starvation and mimosine treatments has been recommended because cell death in these groups was very similar to the control group.