49 INTRA-GENUS CLONED EMBRYOS PRODUCTION AND PREGNANCIES DERIVED FROM LEOPARD CAT NUCLEI AND DOMESTIC CAT ENUCLEATED EGGS

Abstract

The leopard cat (Prionailurus bengalensis), a member of the felidae family, is currently listed as threatened by the Ministry of Environment in South Korea. In exotic or endangered species, the lack of oocytes and recipients precludes the use of traditional somatic cell nuclear transfer (NT), and an approach such as intragenus NT may be the only alternative for producing embryos and offspring. In the present study, we used the leopard cat (LC) as a somatic cell donor to evaluate the in vivo developmental competence, after transfer into domestic cat recipients, of cloned embryos produced by the fusion of LC fibroblast cell nuclei with domestic cat cytoplasts. A total of 412 enucleated domestic cat oocytes were reconstructed with either male (Treatment A) or female (Treatment B) adult LC fibroblasts. There was no significant difference in fusion rate (60.4 vs. 56.9%) between Treatment A and B. Of the fused couplets, the cleavage and blastocyst developmental rate in Treatment A were greater than those in Treatment B (69.5 vs. 60.9%; 8.3 vs. 7.8%; P < 0.05). In treatment A, in vivo developmental studies at 30-45 days postimplantation demonstrated 4.8% (21/435) of reconstructed embryos (n = 435) had entered into the uterine lining of recipients, but only 1.4% (6/435) formed fetuses. However, all of the reconstructed embryos failed to develop to term (65 days). Microsatellite analyses confirmed that the nuclear genome of the cloned fetuses were LC in origin.