Effects of maturational age of recipient oocytes and activation conditions on the development of porcine fetal fibroblast nuclear transfer embryos

Abstract

This study was conducted to evaluate the nuclear remodeling patterns and the developmental potential of porcine fetal fibroblast nuclear transfer embryos (NTs) following the maturational age of recipient oocytes and activation conditions. Donor cells were transferred into the enucleated oocytes that were matured for 36 or 44 h. Electrofused embryos were cultured in PZM-3 for 6 days without activation treatment (EF group). Some of these embryos were additionally activated by electric stimulus (ES; EF + ES group) or a combination of ES and DMAP (EF + ES + D group) before culture. The reconstituted embryos were fixed 2.5 h after fusion to evaluate the nuclear remodeling patterns. The nuclear remodeling pattern of NTs reconstituted with 44 h-matured recipients showed a tendency to form a pronucleus-like structure, while that of NTs reconstituted with 36 h-matured recipients showed a tendency to undergo a premature chromosome condensation (PCC) and form one set of chromatin clump. In EF + ES + D group, blastocyst development was significantly increased regardless of maturational age of recipient oocytes ( P < 0.05). The result indicates that additional activation treatment is necessary to induce the activation of embryos reconstituted with 36 h-matured recipients, and treatment with the combination of electrical stimuli and DMAP could enhance the blastocyst formation rate of porcine NTs reconstituted with both 36 h- and 44 h-matured recipient oocytes.