Dimethyl sulfoxide and glycerol as cryoprotectant agents of stallion semen: Effects on blastocyst rates following intracytoplasmic sperm injection of IVM equine oocytes

Nancy L. Cook, Keith R. Masterson, David Battaglia, Rick Beck, Elizabeth S. Metcalf

Research output: Contribution to journalArticle

Abstract

Numerous variables affect in vitro blastocyst development following intracytoplasmic sperm injection (ICSI). The paternal factor is affected by initial semen quality, processing techniques and final selection of individual spermatozoon for injection. This study investigated whether there was an effect of sperm cryoprotectant agent (CPA) on equine in vitro blastocyst production, and reviews recent developments examining how processing equine semen affects ICSI outcomes. Single ejaculates from five stallions were collected and processed in a freezing extender containing either 1 M dimethyl sulfoxide (DMSO) or 3.5% glycerol. Immature equine oocytes were obtained from ovarian follicles of mares during diestrus by transvaginal aspiration (n = 128). After in vitro maturation, MII oocytes (n = 90) were fertilised by ICSI with thawed stallion spermatozoa (n = 45 in both the DMSO and glycerol groups). The embryo cleavage rate was greater in the DMSO than glycerol group (73.3% vs 46.7% respectively; P = 0.0098), but the blastocyst development rate per fertilised oocyte was similar between the two groups (28.9% vs 15.6% respectively; P = 0.128), as was the blastocyst production rate per cleaved embryo (39.4% vs 33.3% respectively; P = 0.653). In this study, cryopreservation of equine spermatozoa in 1 M DMSO was correlated with significantly higher cleavage rates in IVM oocytes fertilised by ICSI compared with spermatozoa cryopreserved using 3.5% glycerol. Although not statistically significant in this small number of stallions, increased blastocyst production and individual stallion variability was observed among CPA treatments. This warrants further critical examination of cryoprotectants used in equine sperm subpopulations used for ICSI in a larger number of stallions.

Original languageEnglish (US)
JournalReproduction, Fertility and Development
DOIs
StateAccepted/In press - Jan 1 2019

Fingerprint

intracytoplasmic sperm injection
Intracytoplasmic Sperm Injections
Blastocyst
cryoprotectants
stallions
dimethyl sulfoxide
Dimethyl Sulfoxide
Semen
blastocyst
Glycerol
Horses
Oocytes
Spermatozoa
glycerol
semen
oocytes
spermatozoa
horses
embryo (animal)
Embryonic Structures

Keywords

  • assisted reproductive technology
  • cleavage
  • IVF

ASJC Scopus subject areas

  • Biotechnology
  • Reproductive Medicine
  • Animal Science and Zoology
  • Molecular Biology
  • Genetics
  • Endocrinology
  • Developmental Biology

Cite this

Dimethyl sulfoxide and glycerol as cryoprotectant agents of stallion semen : Effects on blastocyst rates following intracytoplasmic sperm injection of IVM equine oocytes. / Cook, Nancy L.; Masterson, Keith R.; Battaglia, David; Beck, Rick; Metcalf, Elizabeth S.

In: Reproduction, Fertility and Development, 01.01.2019.

Research output: Contribution to journalArticle

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abstract = "Numerous variables affect in vitro blastocyst development following intracytoplasmic sperm injection (ICSI). The paternal factor is affected by initial semen quality, processing techniques and final selection of individual spermatozoon for injection. This study investigated whether there was an effect of sperm cryoprotectant agent (CPA) on equine in vitro blastocyst production, and reviews recent developments examining how processing equine semen affects ICSI outcomes. Single ejaculates from five stallions were collected and processed in a freezing extender containing either 1 M dimethyl sulfoxide (DMSO) or 3.5{\%} glycerol. Immature equine oocytes were obtained from ovarian follicles of mares during diestrus by transvaginal aspiration (n = 128). After in vitro maturation, MII oocytes (n = 90) were fertilised by ICSI with thawed stallion spermatozoa (n = 45 in both the DMSO and glycerol groups). The embryo cleavage rate was greater in the DMSO than glycerol group (73.3{\%} vs 46.7{\%} respectively; P = 0.0098), but the blastocyst development rate per fertilised oocyte was similar between the two groups (28.9{\%} vs 15.6{\%} respectively; P = 0.128), as was the blastocyst production rate per cleaved embryo (39.4{\%} vs 33.3{\%} respectively; P = 0.653). In this study, cryopreservation of equine spermatozoa in 1 M DMSO was correlated with significantly higher cleavage rates in IVM oocytes fertilised by ICSI compared with spermatozoa cryopreserved using 3.5{\%} glycerol. Although not statistically significant in this small number of stallions, increased blastocyst production and individual stallion variability was observed among CPA treatments. This warrants further critical examination of cryoprotectants used in equine sperm subpopulations used for ICSI in a larger number of stallions.",
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