Back to Reference Library
veterinary
2019
Case Report

N-acetyl cysteine restores the fertility of vitrified-warmed mouse oocytes derived through ultrasuperovulation.

Authors: Mukunoki Ayumi, Takeo Toru, Nakagata Naomi

Journal: PloS one

Summary

# Editorial Summary Cryopreservation offers valuable opportunities for genetic preservation and fertility management in animals, yet the process causes significant damage to oocytes—particularly hardening of the zona pellucida (the egg's protective outer layer)—which compromises fertilisation rates. This mouse study compared two superovulation protocols and tested whether N-acetyl cysteine (NAC), an antioxidant known to restore thiol chemistry, could reverse vitrification-induced zona damage. The ultrasuperovulation protocol combining inhibin antiserum with equine chorionic gonadotropin produced substantially higher oocyte yields than conventional eCG alone, with comparable post-thaw survival rates; critically, post-warming NAC treatment (0.5 mM) increased fertilisation rates by restoring free thiol levels in the zona and cleaving the abnormal disulfide bonds that accumulate during freezing, allowing normal zona expansion and developmental competence. Embryos generated from NAC-treated oocytes developed into healthy offspring following transfer, demonstrating full restoration of reproductive potential. For equine practitioners managing breeding programmes or genetic resource banking, these findings suggest that combining optimised superovulation techniques with targeted antioxidant treatment of thawed oocytes could substantially improve success rates in assisted reproductive technologies, though further investigation in equine species would be warranted before clinical application.

Read the full abstract on PubMed

Practical Takeaways

  • Not applicable to equine practice—this study used mouse oocytes and reproductive biotechnology techniques not routinely applied to horses
  • Despite equine chorionic gonadotropin (eCG) being mentioned as a treatment agent, the research focuses on laboratory fertility preservation in rodents, not equine clinical applications
  • Results may inform future equine assisted reproductive technology development, but direct clinical translation to horse breeding is not established

Key Findings

  • Ultrasuperovulation with IAS + eCG increased cryopreserved oocyte yield compared to conventional eCG superovulation while maintaining equivalent survival rates
  • N-acetyl cysteine (0.5 mM) increased fertilization rates of vitrified-warmed oocytes by 0.5 mM treatment
  • N-acetyl cysteine restored free thiol levels in zona pellucida and promoted zona expansion by cleaving disulfide bonds
  • Two-cell embryos treated with N-acetyl cysteine developed into normal pups following embryo transfer

Conditions Studied

oocyte cryopreservationsubfertility from vitrificationzona pellucida hardening

Related References

Superovulation using the combined administration of inhibin antiserum and equine chorionic gonadotropin increases the number of ovulated oocytes in C57BL/6 female mice.

Takeo Toru, Nakagata Naomi(2015)PloS one

Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen-Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status.

Gimeno Brenda Florencia, Bariani María Victoria, Laiz-Quiroga Lucía, Martínez-León Eduardo, Von-Meyeren Micaela, Rey Osvaldo, Mutto Adrián Ángel, Osycka-Salut Claudia Elena(2021)Animals : an open access journal from MDPI

New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification.

Angel-Velez Daniel, De Coster Tine, Azari-Dolatabad Nima, Fernandez-Montoro Andrea, Benedetti Camilla, Bogado Pascottini Osvaldo, Woelders Henri, Van Soom Ann, Smits Katrien(2021)Animals : an open access journal from MDPI

Synchronization of the ovulation and copulation timings increased the number of in vivo fertilized oocytes in superovulated female mice.

Nakao Satohiro, Ito Kotono, Sugahara Chihiro, Watanabe Hitomi, Kondoh Gen, Nakagata Naomi, Takeo Toru(2023)PloS one

Effect of Using Two Cryopreservation Methods on Viability and Fertility of Frozen Stallion Sperm.

Maziero Rosiara Rosaria Dias, Guaitolini Carlos Renato de Freitas, Guasti Priscilla Nascimento, Monteiro Gabriel Augusto, Martin Ian, Silva Juliano Pianowski Marques da, Crespilho André Maciel, Papa Frederico Ozanam(2019)Journal of equine veterinary science