Production of sexed bovine embryos in vitro can be improved by selection of sperm treatment and co-culture system

Reprod Domest Anim. 2021 Jun;56(6):864-871. doi: 10.1111/rda.13926. Epub 2021 Apr 4.

Abstract

The study investigated the effects of sperm sorting, capacitation treatment and co-cultivation on sexed bovine in vitro embryo production. The effect of treatment and co-culture on production of embryos of the preferred sex from unsorted sperm was also studied. Sperm from five breeding bulls was used for fertilization of mature oocytes as follows: Experiment 1, sorted and unsorted sperm (bulls A-E) treated only with heparin in standard co-cultures; Experiment 2, sorted sperm (bulls A-E) treated with heparin-PHE (penicillamine, hypotaurine, and epinephrine) or heparin-caffeine in drop co-cultures; and Experiment 3, unsorted sperm (bull E) treated with either heparin-PHE or heparin-caffeine in both standard and drop co-cultures. In all bulls, treatment with heparin resulted in significantly (p < .05) reduced cleavage and blastocyst rates from sorted sperm, as compared with those from unsorted sperm. In bulls A, B, D and E, treatment of sorted sperm with heparin-PHE in drops significantly increased the blastocyst rate (p < .05). In unsorted sperm of bull E, heparin-PHE treatment in drops resulted in the XX/XY sex ratio inverse to that obtained by heparin-caffeine treatment in standard co-cultures (32.3%/67.7% and 66.7%/33.3%, respectively). In conclusion, the treatment of sorted sperm with heparin-PHE in modified drop co-cultures can be recommended for production of in vitro sexed embryos. The use of unsorted sperm for production of embryos of the preferred sex by selected capacitation treatment and co-culture can be the method of choice in bulls with low IVF yields from sorted sperm.

Keywords: IVF; breeding bulls; capacitation treatment; sexed embryos; sorting; sperm.

MeSH terms

  • Animals
  • Cattle
  • Coculture Techniques / methods
  • Coculture Techniques / veterinary*
  • Embryo Culture Techniques / veterinary
  • Epinephrine / pharmacology
  • Female
  • Fertilization in Vitro / methods
  • Fertilization in Vitro / veterinary
  • Heparin / pharmacology
  • Male
  • Oocytes
  • Penicillamine / pharmacology
  • Sex Preselection / methods
  • Sex Preselection / veterinary*
  • Spermatozoa / drug effects*
  • Taurine / analogs & derivatives
  • Taurine / pharmacology

Substances

  • Taurine
  • hypotaurine
  • Heparin
  • Penicillamine
  • Epinephrine