Establishment of bovine Fetal Fibroblasts Line for Production of Cloned Calves in Korean Native Cattle: The Effects of Culture Period and Various Cell Size on the Efficiency of Nuclear Transfer

복제 한우 생산을 위한 Bovine Fetal Fibroblasts의 이용에 관한 연구: 공여핵원의 배양기간 및 세포 크기가 핵이식의 효율에 미치는 영향

The development potential of bovine somatic cells was evaluated using nuclear transfer. A single donor cell derived from fetus of HanWoo(Korean Native Cattle) was selected and deposited into perivitelline space of each enucleated oocyte before electrical fusion and activation. Nuclei of donor cells starved for 7 days (37%) tended to support the development of reconstitute embryo the blastocyst stage better than those of donor cells starved 3, 14 and 30 days. The cleavage rate was significantly lower(P<0.05) in reconstitute embryos derived from large size donor cells(51.2%), than those from small medium size donor cells(76.6 and 73.5, respectively). The developmental rate to blastocyst of reconstructed embryos from medium size donor cells was higher than those from small and medium size donor cells. This study demonstrates that an appropriate culture period for induction into quiescent stage and the size of donor cells effect on the efficiency of nuclear transfer using cultured bovine cells.