Developmental Efficiency of Bovine Embryos Cloned with Fetal Fibroblast Arrested at G0/G1 Phase

The study evaluated the effect of donor cell treatments for G0/Gl synchronization and the donor ceil type on development and incidence of apoptosis in cloned cattle embryos. Primary cultures were established from a female fetus on day 50 of gestation and adult ear skin biopsies. Cells were randomly allocated into 3 experimental treatment groups after 6~8 passages. Group 1 (Confluent), cells were cultured in DMEM supplemented with 10% FBS until 90% confluent. Group 2 (Serum-starvation), cells were cultured in DMEM Supplemented With 0.5% FBS for 5 days. Group 3 (Roscovitine), Cells were cultured in DMEM supplemented with 10% FBS and 30 $\mu$M Roscovitine for 12 h. Cell cycle and apoptosis were analyzed using flow cytometry after labelling with DAPI and YO-PRO-1. At 19 h post-maturation (hpm), enucleated oocytes were reconstructed with donor cells and fused by a single DC pulse (1.6 kV/cm, 60 $\mu$sec). (중략)