Theriogenology2005; 63(9); 2372-2381; doi: 10.1016/j.theriogenology.2004.05.032

Effect of seminal plasma on the cryopreservation of equine spermatozoa.

Abstract: Seminal plasma is generally removed from equine spermatozoa prior to cryopreservation. Two experiments were designed to determine if adding seminal plasma back to spermatozoa, prior to cryopreservation, would benefit the spermatozoa. Experiment 1 determined if different concentrations of seminal plasma affected post-thaw sperm motility, viability and acrosomal integrity of frozen/thawed stallion spermatozoa. Semen was washed through 15% Percoll to remove seminal plasma and spermatozoa resuspended to 350 x 10(6)sperm/mL in a clear Hepes buffered diluent containing either 0, 5, 10, 20, 40 or 80% seminal plasma for 15 min, prior to being diluted to a final concentration of 50 x 10(6)sperm/mL in a Lactose-EDTA freezing diluent and cryopreserved. Sperm motility was analyzed at 10 and 90 min after thawing, while sperm viability and acrosomal integrity were analyzed 20 min after thawing. Seminal plasma did not affect sperm motility, viability or acrosomal integrity (P>0.05). Experiment 2 tested the main affects of seminal plasma level (5 or 20%), incubation temperature (5 or 20 degrees C) and incubation time (2, 4 or 6 h) prior to cryopreservation. In this experiment, spermatozoa were incubated with 5 or 20% seminal plasma for up to 6h at either 5 or 20 degrees C prior to cryopreservation in a skim milk, egg yolk freezing extender. Samples cooled immediately to 5 degrees C, prior to freezing had higher percentages of progressively motile spermatozoa than treatments incubated at 20 degrees C (31 versus 25%, respectively; P<0.05), when analyzed 10 min after thawing. At 90 min post-thaw, total motility was higher for samples incubated at 5 degrees C (42%) compared to 20 degrees C (35%; P<0.05). In addition, samples containing 5% seminal plasma had higher percentages of total and progressively motile spermatozoa (45 and 15%) than samples exposed to 20% seminal plasma (33 and 9%; P<0.05). In conclusion, although the short-term exposure of sperm to seminal plasma had no significant effect on the motility of cryopreserved equine spermatozoa, prolonged exposure to seminal plasma, prior to cryopreservation, was deleterious.
Publication Date: 2005-05-25 PubMed ID: 15910920DOI: 10.1016/j.theriogenology.2004.05.032Google Scholar: Lookup
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  • Journal Article

Summary

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The research investigates whether adding seminal plasma back to horse sperm before freezing would have any benefits. However, the results show that seminal plasma does not significantly impact the quality of frozen sperm, and that prolonged exposure to seminal plasma before freezing may even be harmful.

Objective of the Research

  • This study aims to determine whether adding seminal plasma back to equine sperm prior to cryopreservation (a process of freezing biological construct) would bring any benefit to the sperm’s quality.

First Experiment

  • Firstly, semen was washed to remove seminal plasma and was then resuspended at a given concentration.
  • After this, seminal plasma was reintroduced to the suspension at varying percentages, and the suspension was cryopreserved.
  • The post-thaw sperm motility (movement abilities), viability (ability to function), and acrosomal integrity (structural soundness of a part of the sperm cell) were then analyzed.
  • The results showed that the addition of seminal plasma did not affect the quality of the sperm.

Second Experiment

  • The main effects of seminal plasma level, incubation temperature, and incubation time prior to cryopreservation were tested in the second experiment.
  • In this experiment, temperatures and time of incubation varied and the samples were again cryopreserved.
  • Again, sperm quality was analyzed after thawing.
  • The results showed that samples incubated at a lower temperature showed higher sperm motility than those incubated at a higher temperature. In addition, samples containing less seminal plasma demonstrated increased motility than those with a higher concentration of seminal plasma.

Conclusion

  • The study concludes that although short-term exposure to seminal plasma doesn’t significantly impact the quality of cryopreserved equine sperm, prolonged exposure before freezing may be harmful.
  • Thus, it might be better to avoid adding back seminal plasma to horse spermatozoa just before cryopreservation considering it doesn’t benefit and could potentially damage the sperm quality.

Cite This Article

APA
Moore AI, Squires EL, Graham JK. (2005). Effect of seminal plasma on the cryopreservation of equine spermatozoa. Theriogenology, 63(9), 2372-2381. https://doi.org/10.1016/j.theriogenology.2004.05.032

Publication

ISSN: 0093-691X
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 63
Issue: 9
Pages: 2372-2381

Researcher Affiliations

Moore, A I
  • Animal Reproduction and Biotechnology Laboratory, Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80521, USA.
Squires, E L
    Graham, J K

      MeSH Terms

      • Animals
      • Cell Survival
      • Cryopreservation / veterinary
      • Horses
      • Male
      • Semen / physiology
      • Semen Preservation / methods
      • Semen Preservation / veterinary
      • Sperm Motility
      • Spermatozoa / physiology

      Citations

      This article has been cited 16 times.
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