Investigation of the effects of different cell synchronization methods on early and late passage number sheep and bovine cells

The cell cycle stage of donor cells is an important factor influencing developmental ability of nuclear transfer embryos. G0 or G1 stages are considered to be the most suitable stage for nuclear transfer. In this study, the granulosa cells from cattle and sheep were analyzed low (5th) and high (15th) passages. Experimental groups consiste actively dividing cells, contact inhibition (%10 FBS) for 24 and 72 hours, cells starvation (%0.5 FBS) for 24 and 72 hours. Cell cycle phases ratios were determined after cells were stained with propidium iodide, cell viability/apoptosis/necrosis cell ratios were determined after cells were stained with propidium iodide and Annexin V-FITC by flow cytometry In the treatment groups, it was observed that the G0/G1 rate was between %78.69-87.5 in confluent cells for 24/72 hours, and the G0/G1 rate was between %83.02-%96.16 in serum starvation cells for 24/72 hours found to be higher than the control group (%68.22-%76.97) (P<0.05). According to the analysis findings, it was determined that the total rate of early and late apoptotic cells varied between %1.17-%4.42, and necrosis cells varied between %1.41-%6.46. Except for sheep high (15th) passage, the best synchronization was obtained from serum starvation experimental groups. There was no statistical difference between the sheep higher (15th) passage application groups (P<0.05).