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

Zygote. 2019 Jun;27(3):137-142. doi: 10.1017/S0967199419000029. Epub 2019 Apr 30.

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.

Keywords: Cattle; Cytoplast; DNA damage; Livestock; Nuclear transplantation.

MeSH terms

  • Animals
  • Antibiotics, Antineoplastic / pharmacology
  • Blastocyst / cytology
  • Blastocyst / drug effects*
  • Blastocyst / metabolism
  • Cattle
  • Cell Nucleus / metabolism*
  • Cloning, Organism / methods*
  • Cloning, Organism / veterinary
  • Embryo Transfer
  • Embryonic Development
  • Female
  • Mitomycin / pharmacology*
  • Nuclear Transfer Techniques / standards*
  • Nuclear Transfer Techniques / veterinary
  • Oocytes / cytology
  • Oocytes / drug effects*
  • Oocytes / metabolism
  • Parthenogenesis
  • Pregnancy

Substances

  • Antibiotics, Antineoplastic
  • Mitomycin