Production of somatic cell nuclear transfer embryos using in vitro-grown and in vitro-matured oocytes in rabbits

Zygote. 2015 Aug;23(4):494-500. doi: 10.1017/S0967199414000082. Epub 2014 Mar 26.

Abstract

We examined growing oocytes collected from follicles remaining in superovulated rabbit ovaries, that were grown (in vitro growth, IVG) and matured (in vitro maturation, IVM) in vitro. We produced somatic cell nuclear transfer (SCNT) embryos using the mature oocytes and examined whether these embryos have the ability to develop to the blastocyst stage. In addition, we examined the effects of trichostatin A (TSA), a histone deacetylase inhibitor (HDACi), on the developmental competence of SCNT embryos derived from IVG-IVM oocytes. After growth for 7 days and maturation for 14-16 h in vitro, the growing oocytes reached the metaphase II stage (51.4%). After SCNT, these reconstructed embryos reached the blastocyst stage (20%). Furthermore, the rate of development to the blastocyst stage and the number of cells in the blastocysts in SCNT embryos derived from IVG-IVM oocytes were significantly higher for TSA-treated embryos compared with TSA-untreated embryos (40.6 versus 21.4% and 353.1 ± 59.1 versus 202.5 ± 54.6, P < 0.05). These results indicate that rabbit SCNT embryos using IVG-IVM oocytes have the developmental competence to reach the blastocyst stage.

Keywords: Follicle; In vitro growth; Nuclear transfer; Rabbit; Trichostatin A.

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Blastocyst / cytology
  • Blastocyst / drug effects*
  • Blastocyst / physiology
  • Embryo, Mammalian / drug effects
  • Female
  • Histone Deacetylase Inhibitors / pharmacology
  • Hydroxamic Acids / pharmacology*
  • In Vitro Oocyte Maturation Techniques / methods*
  • Metaphase
  • Nuclear Transfer Techniques*
  • Oocytes / cytology
  • Oocytes / physiology
  • Rabbits

Substances

  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • trichostatin A