Bovine somatic cell nuclear transfer using mitomycin C-mediated chemical oocyte enucleation.

ABSTRACT

SummaryChemical oocyte enucleation holds the potential to ease somatic cell nuclear transfer (SCNT), although high enucleation rates remain limited to micromanipulation-based approaches. Therefore, this study aimed to test mitomycin C (MMC) for use in bovine functional chemical oocyte enucleation. Incubation of denuded eggs in 10 µg ml-1 MMC for different periods did not affect most maturation rates (0.5 h 85.78%A, 1.0 h 72.77%B, 1.5 h 83.87%A, and 2.0 h 82.05%A) in comparison with non-treated controls (CTL; 85.77%A). Parthenogenetic development arrest by MMC was efficient at cleavage (CTL 72.93%A, 0.5 h 64.92%A,B, 1.0 h 60.39%B,C, 1.5 h 66.35%A,B, and 2.0 h 53.84%C) and blastocyst stages (CTL 33.94%A, 0.5 h 7.58%B, 1.0 h 2.47%C, 1.5 h 0.46%C, and 2.0 h 0.51%C). Blastocysts were obtained after nuclear transfer (NT) using MMC enucleation [NT(MMC) 4.54%B] but at lower rates than for the SCNT control [NT(CTL) 26.31%A]. The removal of the meiotic spindle after MMC incubation fully restored SCNT blastocyst development [NT(MMC+SR) 24.74%A]. Early pregnancies were obtained by the transfer of NT(MMC) and NT(MMC+SR) blastocysts to synchronized recipients. In conclusion, MMC leads to functional chemical oocyte enucleation during SCNT and further suggests its potential for application towards technical improvements.