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 language | English (US) |
|---|---|
| Journal | Reproduction, Fertility and Development |
| DOIs | |
| State | Accepted/In press - Jan 1 2019 |
Fingerprint
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 journal › Article
}
TY - JOUR
T1 - Dimethyl sulfoxide and glycerol as cryoprotectant agents of stallion semen
T2 - Effects on blastocyst rates following intracytoplasmic sperm injection of IVM equine oocytes
AU - Cook, Nancy L.
AU - Masterson, Keith R.
AU - Battaglia, David
AU - Beck, Rick
AU - Metcalf, Elizabeth S.
PY - 2019/1/1
Y1 - 2019/1/1
N2 - 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.
AB - 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.
KW - assisted reproductive technology
KW - cleavage
KW - IVF
UR - http://www.scopus.com/inward/record.url?scp=85076374708&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85076374708&partnerID=8YFLogxK
U2 - 10.1071/RD19266
DO - 10.1071/RD19266
M3 - Article
AN - SCOPUS:85076374708
JO - Reproduction, Fertility and Development
JF - Reproduction, Fertility and Development
SN - 1031-3613
ER -