Methyl-beta cyclodextrin and creatine work synergistically under hypoxic conditions to improve the fertilization ability of boar ejaculated sperm

Anim Sci J. 2020 Jan-Dec;91(1):e13493. doi: 10.1111/asj.13493.


Although successful fertilization is completed by only 150 sperm in the pig oviduct, more than 50,000 sperms are required to achieve a fertilization rate of more than 70% by pig in vitro fertilization (IVF). In this study, to improve the efficiency of pig IVF, the effects of hypoxic conditions and treatment with creatine and methyl-beta cyclodextrin (MβCD) on the glycolytic pathway were investigated. Under low O2 conditions, zig-zag motility was strongly induced within 30 min; however, the induction disappeared at 60 min. Although caffeine suppressed zig-zag motility under low O2 conditions, creatine induced and sustained zig-zag motility until 120 min. Additionally, pretreatment with MβCD for 15 min greatly enhanced zig-zag motility via ATP production in sperm incubated with creatine under low O2 conditions. Sperm pretreated with MβCD were used for IVF in medium containing creatine under low O2 conditions. A fertilization rate of approximately 70% was achieved with only 1.0 x 104 sperms/mL, and there were few polyspermic embryos. Therefore, our novel method was beneficial for efficient production of pig embryos in vitro. Moreover, the zig-zag motility may be a novel movement which boar capacitated sperm exhibit in the culture medium.

Keywords: boar sperm; in vitro fertilization; polyspermy; zig-zag motility.

MeSH terms

  • Anaerobiosis / physiology*
  • Animals
  • Creatine / pharmacology*
  • Drug Synergism
  • Ejaculation / physiology
  • Female
  • Fertilization / drug effects*
  • Fertilization / physiology
  • Fertilization in Vitro / methods*
  • Fertilization in Vitro / veterinary*
  • Male
  • Sperm Motility*
  • Sperm-Ovum Interactions / drug effects*
  • Sperm-Ovum Interactions / physiology
  • Spermatozoa / physiology*
  • Swine / physiology*
  • beta-Cyclodextrins / pharmacokinetics*
  • beta-Cyclodextrins / pharmacology


  • beta-Cyclodextrins
  • Creatine