Bovine and Swamp Buffalo Somatic Cell Nuclear Transfer in Thailand

This experiment was studied to examine the in vitro developmental potential of enucleateddomestic cat oocytes reconstructed with skin fibroblasts from female domestic and leopard cats. The cumulus oocyte complex of domestic cat were cultured in IVM medium under 38.5°C, 5%CO2, 5%O2,90%N2 for 24 hours. The matured oocytes were enucleated and individual donor cell was inserted intoperivitelline space of enucleated oocyte. The couples were fused by electrostimulation (2DC, 30 Volt, 30 μsec). The reconstructed embryos were activated by 7% Ethanol for 5 minutes and incubated in 1.25 μg/ml Cytochalasin D and 10 μg/ml Cycloheximide for 5 hours. After activated, the embryos were cultured in Tyrode’s medium + 0.3% BSA under 38.5°C, 5%CO2, 5%O2, 90%N2 for 2 days. Then the8-cells embryos were cultured in Tyrode’s medium + 10%FCS under 38.5°C,5%CO2, 5%O2, 90%N2for 5 days. Parthenogenetic embryos had also performed by the same activation protocol. The cleavagerate of parthenotes and cloned embryos were not different. The proportions of parthenote blastocystsand cloned blastocysts derived from domestic cat fibroblasts (8/56, 14.3% and 7/51, 13.7%respectively) were significantly higher than those blastocysts derived from leopard cat fibroblasts (3/45,6.7%). These results indicated that enucleated domestic cat oocytes could be used as recipient cytoplastfor leopard cat cloning.