RepSox improves viability and regulates gene expression in rhesus monkey-pig interspecies cloned embryos

Biotechnol Lett. 2017 May;39(5):775-783. doi: 10.1007/s10529-017-2308-3. Epub 2017 Feb 28.

Abstract

Objective: To investigate the effect of the small molecule, RepSox, on the expression of developmentally important genes and the pre-implantation development of rhesus monkey-pig interspecies somatic cell nuclear transfer (iSCNT) embryos.

Results: Rhesus monkey cells expressing the monomeric red fluorescent protein 1 which have a normal (42) chromosome complement, were used as donor cells to generate iSCNT embryos. RepSox increased the expression levels of the pluripotency-related genes, Oct4 and Nanog (p < 0.05), but not of Sox2 compared with untreated embryos at the 2-4-cell stage. Expression of the anti-apoptotic gene, Bcl2, and the pro-apoptotic gene Bax was also affected at the 2-4-cell stage. RepSox treatment also increased the immunostaining intensity of Oct4 at the blastocyst stage (p < 0.05). Although the blastocyst developmental rate was higher in the group treated with 25 µM RepSox for 24 h than in the untreated control group (2.4 vs. 1.2%, p > 0.05), this was not significant.

Conclusion: RepSox can improve the developmental potential of rhesus monkey-pig iSCNT embryos by regulating the expression of pluripotency-related genes.

Keywords: Embryos (monkey–pig); Interspecies (monkey–pig) somatic cell nuclear transfer; Pluripotency; RepSox; Rhesus monkey; mRFP1.

MeSH terms

  • Animals
  • Cloning, Organism / methods*
  • Embryo, Mammalian / metabolism*
  • Gene Expression Regulation / drug effects*
  • Induced Pluripotent Stem Cells / metabolism
  • Macaca mulatta
  • Nanog Homeobox Protein / metabolism
  • Nuclear Transfer Techniques*
  • Octamer Transcription Factor-3
  • Oocytes / metabolism
  • Pyrazoles / pharmacology*
  • Pyridines / pharmacology*
  • Swine

Substances

  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3
  • Pyrazoles
  • Pyridines
  • RepSox