Assessment of developmental potential of caprine cloned embryos with ooplasm replenishment under two culture media

The present study was designed to assess the developmental potential of somatic cell nuclear transfer (SCNT) embryos, with and without replenishment of ooplasm into enucleated oocytes, by culturing separately in two culture media. The enucleated oocytes were replenished with exogenous matured ooplasm under replenished nuclear transfer (RNT) method and compared with conventional nuclear transfer (CNT) method without replenishment. The fusion efficiency in RNT group was found to be significantly higher (P < 0.05) than CNT group (59.39 ± 7.36 vs 45.57 ± 3.68%). The completely fused reconstructed oocytes from both groups were cultured separately in research vitro cleave medium (RVCL) and RVCL-Blast medium. The embryonic development of 2 cell, 4 cell, 8-16 cell and 16-32 cell stages in the RNT group was superior to the CNT group regardless of the culture media used (P < 0.05). The embryonic development of the 8-16 cell, 16-32 cell, morula, and blastocyst stages in the RVCL-Blast medium was significantly higher (P < 0.05) than the RVCL alone for both RNT as well as CNT groups. RNT method with RVCL-Blast produced highly significant (P < 0.01) embryonic development for 8-16 cell and 16-32 cell stage when compared to CNT with RVCL. Conclusively, the combination of RNT with RVCL-Blast culture media enabled an overall increase in the embryonic developmental potential.