Effects of trichostatin A on in vitro development and transgene function in somatic cell nuclear transfer embryos derived from transgenic Clawn miniature pig cells

Anim Sci J. 2010 Oct;81(5):558-63. doi: 10.1111/j.1740-0929.2010.00772.x.

Abstract

The present study was carried out to examine the effects of post-activation treatment of trichostatin A (TSA), a histone deacetylase inhibitor, on in vitro development and transgene function of somatic cell nuclear transfer (SCNT) embryos derived from Clawn miniature pig embryonic fibroblast (PEF) transfected with a bacterial endo-β-galactosidase C gene (removal of the α-galactosyl (Gal) epitope). SCNT embryos were incubated with or without TSA (50 or 100 nmol/L) after activation, cultured in vitro and assessed for cleavage, blastocyst formation and transgene function. The rate of blastocyst formation was significantly higher in SCNT embryos treated with 50 nmol/L TSA than that in control (P < 0.05), whereas the rate of cleavage and cell number of blastocyst did not differ. Following labelling with fluorescein isothiocyanate-labelled BS-I-B(4) isolectin, the intensity of fluorescence observed on cell-surface was dramatically reduced in transgenic SCNT blastocyst in comparison with non-transgenic SCNT blastocyst. However, the reduction of α-Gal epitope expression in transgenic SCNT blastocyst was not affected by TSA treatment. The results of this study showed that post-activation treatment with 50 nmol/L TSA is effective to improve in vitro developmental capacity of transgenic SCNT miniature pig embryos without the modification of transgene function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Embryo, Mammalian / drug effects
  • Histone Deacetylase Inhibitors / pharmacology*
  • Hydroxamic Acids / pharmacology*
  • Nuclear Transfer Techniques / veterinary*
  • Swine
  • Swine, Miniature / embryology*
  • Transgenes / drug effects*

Substances

  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • trichostatin A