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Methods in molecular biology (Clifton, N.J.)2020; 2180; 365-377; doi: 10.1007/978-1-0716-0783-1_15

Cryopreservation of Semen from Domestic Livestock: Bovine, Equine, and Porcine Sperm.

Abstract: In modern livestock breeding, cryopreserved semen is routinely used for artificial insemination. Sperm cryopreservation allows for long-term storage of insemination doses and secures reproduction at a desired time point. In order to cryopreserve semen, it needs to be carefully processed to preserve its vital functions after thawing. In this chapter, we describe the processes involved in cryopreservation of bull, stallion, and boar sperm. These include preparation of diluents, dilution of sperm in primary and freezing extender, slow cooling from room temperature to 5 °C, packaging of insemination doses in straws, freezing at a defined cooling rate in liquid nitrogen vapor, cryogenic storage, and thawing. Two-step dilution approaches, with commonly used diluents, are presented, namely, TRIS-egg yolk (TEY) extender for bull sperm, skim milk (INRA-82) extender for stallion sperm, and lactose-egg yolk (LEY) extender for boar sperm. Furthermore, simple methods are presented for cooling and freezing of sperm at defined cooling rates.
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Publication Date: 2020-08-17 PubMed ID: 32797421DOI: 10.1007/978-1-0716-0783-1_15Google 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.

This study describes the methodology of cryopreserving sperm from three domestic livestock species: boars, stallions, and bulls. The researchers outline key protocols including preparing diluents and dilution of sperm in extender, progressive cooling, packaging in straws, freezing in liquid nitrogen, cryogenic storage, and thawing.

Methodology

  • The process starts with the preparation of diluents that are used to extend the life span of the semen upon freezing and thawing.
  • Next, the sperm is diluted in primary and freezing extenders. These extenders vary based on the species: for example, bull sperm is processed in a TRIS-egg yolk (TEY) extender, stallion sperm is processed in skim milk (INRA-82) extender, and boar sperm is processed in a lactose-egg yolk (LEY) extender.
  • The diluted sperm is then cooled slowly from room temperature to 5°C. This gradual cooling allows the semen to undergo physiological changes necessary to withstand the freezing and thawing process.
  • The cooled semen is then packaged into straws for individual servings, ready for artificial insemination.

Freezing and storing process

  • The packaged semen is then frozen at a defined cooling rate using liquid nitrogen vapor. Controlling the freezing rate is crucial as too rapid or too slow freezing can harm the sperm and lessen its ability to fertilize an egg.
  • Once frozen, the semen is stored in cryogenic conditions until needed. Using cryogenic storage allows for the preservation of genetic material for extended periods, protecting it from deterioration.

Thawing

  • When the time comes for the frozen semen to be used in artificial insemination, a specific thawing process is necessary to restore its fertility potential.
  • The researchers present simple methods for cooling and freezing sperm at these defined cooling rates to maximize post-thaw vitality.

Through this research, greater insight is offered into the intricate process of livestock sperm cryopreservation, an important tool in modern breeding practices.

Cite This Article

APA
Oldenhof H, Wolkers WF, Sieme H. (2020). Cryopreservation of Semen from Domestic Livestock: Bovine, Equine, and Porcine Sperm. Methods Mol Biol, 2180, 365-377. https://doi.org/10.1007/978-1-0716-0783-1_15

Publication

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
NlmUniqueID: 9214969
Country: United States
Language: English
Volume: 2180
Pages: 365-377

Researcher Affiliations

Oldenhof, Harriëtte
  • Unit for Reproductive Medicine-Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany. harriette.oldenhof@tiho-hannover.de.
Wolkers, Willem F
  • Unit for Reproductive Medicine-Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany.
  • Biostabilization Laboratory-Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, University of Veterinary Medicine Hannover, Hannover, Germany.
Sieme, Harald
  • Unit for Reproductive Medicine-Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany.

MeSH Terms

  • Animals
  • Cattle
  • Cryopreservation / methods
  • Cryopreservation / veterinary
  • Cryoprotective Agents / pharmacology
  • Horses
  • Male
  • Semen / drug effects
  • Semen / physiology
  • Semen Preservation / methods
  • Semen Preservation / veterinary
  • Swine

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Citations

This article has been cited 14 times.
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