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Home / Equine Research Bank / Reprod Domest Anim / Prolonged incubation of frozen-thawed equine spermatozoa for...
Reproduction in domestic animals = Zuchthygiene2024; 59 Suppl 3; e14593; doi: 10.1111/rda.14593

Prolonged incubation of frozen-thawed equine spermatozoa for in vitro fertilization: A preliminary study using low temperature and INRA96 medium.

Abstract: A protocol for conventional in vitro fertilization (IVF) in horses using fresh semen has been described, using a prolonged incubation in FERT-TALP medium (22 h) at 38.2°C in the presence of penicillamine, hypotaurine and epinephrine (PHE). Our work aimed to develop a protocol that maintains quality parameters in frozen-thawed equine spermatozoa incubated for 22 h in the presence of PHE using different media (FERT-TALP and INRA96) and incubation temperatures (30 and 38.2°C). Twelve frozen ejaculates from four stallions were thawed and then incubated in either FERT-TALP or INRA96 with PHE at 30 or 38.2°C for 22 h. Following incubation, total motility (TM), progressive motility (PM), viability and acrosome integrity were evaluated. The results showed that TM was significantly higher (p < .001) at 30°C in both media, while PM was higher for INRA96 at 30°C compared to 38°C (p < .05). Moreover, INRA96 at 30°C exhibited higher sperm viability and acrosome integrity (p < .001) compared to the other experimental groups. These preliminary results suggest that incubating thawed equine spermatozoa at 30°C with PHE in INRA96 successfully maintains motility, viability and acrosome integrity in equine spermatozoa, indicating its potential use for conventional equine IVF.
© 2024 The Authors. Reproduction in Domestic Animals published by Wiley‐VCH GmbH.
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Publication Date: 2024-10-14 PubMed ID: 39396854DOI: 10.1111/rda.14593Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research attempts to develop a method for keeping frozen-thawed horse sperm viable during in vitro fertilization, specifically using a prolonged 22-hour incubation in the presence of certain compounds at different temperatures and media. They found that the optimal conditions were using INRA96 medium at 30°C, which maintained sperm motility, viability, and acrosome integrity.

Research Purpose

  • The aim of this study was to develop a new protocol that would maintain key quality parameters in frozen-thawed equine spermatozoa during a prolonged incubation period. The researchers were specifically interested in the effects of different incubation temperatures and media on the sperm.

Methodology

  • The researchers thawed and incubated twelve frozen ejaculates from four stallions in either FERT-TALP or INRA96 media. These media contained compounds known as PHE (penicillamine, hypotaurine, and epinephrine).
  • They then incubated these samples at two different temperatures, 30°C and 38.2°C, for 22 hours.
  • The team evaluated the total motility, progressive motility, viability and acrosome integrity of the sperm after incubation.

Findings

  • The researchers found that the total motility of the sperm was significantly higher when incubated at 30°C in both types of media.
  • For progressive motility, the best results were achieved with INRA96 medium at 30°C, compared to 38°C.
  • The study also found that sperm incubated in INRA96 medium at 30°C had higher viability and acrosome integrity compared to the other experimental groups.

Conclusion

  • The preliminary results suggest that incubating thawed equine sperm at 30°C in INRA96 media with PHE successfully maintains the motility, viability and acrosome integrity of equine sperm. These are important factors for successful in vitro fertilization.
  • The findings indicate the potential use of this method for conventional equine IVF, though further research is necessary to confirm the full effectiveness and applicability of the procedure.

Cite This Article

APA
Luis-Calero M, Muñoz-García CC, Fernández-Hernández P, Macías-García B, González-Fernández L. (2024). Prolonged incubation of frozen-thawed equine spermatozoa for in vitro fertilization: A preliminary study using low temperature and INRA96 medium. Reprod Domest Anim, 59 Suppl 3, e14593. https://doi.org/10.1111/rda.14593

Publication

Reproduction in domestic animals = Zuchthygiene
ISSN: 1439-0531
NlmUniqueID: 9015668
Country: Germany
Language: English
Volume: 59 Suppl 3
Pages: e14593

Researcher Affiliations

Luis-Calero, Marcos
  • Departamento de Medicina Animal, Grupo de Investigación Medicina Interna Veterinaria (MINVET), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
Muñoz-García, Carmen C
  • Departamento de Medicina Animal, Grupo de Investigación Medicina Interna Veterinaria (MINVET), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
Fernández-Hernández, Pablo
  • Departamento de Medicina Animal, Grupo de Investigación Medicina Interna Veterinaria (MINVET), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
Macías-García, Beatriz
  • Departamento de Medicina Animal, Grupo de Investigación Medicina Interna Veterinaria (MINVET), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
González-Fernández, Lauro
  • Departamento de Bioquímica y Biología Molecular y Genética, Grupo de Investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.

MeSH Terms

  • Animals
  • Horses / physiology
  • Male
  • Semen Preservation / veterinary
  • Semen Preservation / methods
  • Fertilization in Vitro / veterinary
  • Cryopreservation / veterinary
  • Spermatozoa / physiology
  • Sperm Motility
  • Acrosome
  • Culture Media
  • Cold Temperature
  • Epinephrine / pharmacology
  • Cell Survival

Grant Funding

  • IB20078, IB20005 and PI-0152-22 / Junta de Extremadura
  • RC1 / Universidad de Extremadura and Banco Santander
  • IB20078 and IB20005 / European Regional Development Fund
  • PID2020-112723RB-I00 / Agencia Estatal de Investigación

References

This article includes 9 references
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