A Double Nuclear Transfer Technique for Cloning Pigs

The first round of double nuclear transfer (NT) procedure includes the following steps: transfer of somatic cell nuclei into enucleated recipient oocytes, fusion, activation, and culture of reconstructed oocytes. The next day, a second round of NT is performed by removing karyoplasts from 1-d-old NT embryos and transferring them into in vivo-derived zygotes from which the two pronuclei have been removed. Couplets are then fused using an electrical pulse and transferred into synchronized recipient gilts. This system, which uses fertilized oocytes as cytoplast recipients, bypasses the inefficiencies of artificial activation procedures, and may promote more successful development.