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Acta veterinaria Scandinavica2012; 54(1); 72; doi: 10.1186/1751-0147-54-72

The effect of two pre-cryopreservation single layer colloidal centrifugation protocols in combination with different freezing extenders on the fragmentation dynamics of thawed equine sperm DNA.

Abstract: Variability among stallions in terms of semen cryopreservation quality renders it difficult to arrive at a standardized cryopreservation method. Different extenders and processing techniques (such us colloidal centrifugation) are used in order to optimize post-thaw sperm quality. Sperm chromatin integrity analysis is an effective tool for assessing such quality. The aim of the present study was to compare the effect of two single layer colloidal centrifugation protocols (prior to cryopreservation) in combination with three commercial freezing extenders on the post-thaw chromatin integrity of equine sperm samples at different post-thaw incubation (37°C) times (i.e., their DNA fragmentation dynamics). Results: Post-thaw DNA fragmentation levels in semen samples subjected to either of the colloidal centrifugation protocols were significantly lower (p<0.05) immediately after thawing and after 4 h of incubation at 37°C compared to samples that underwent standard (control) centrifugation. The use of InraFreeze® extender was associated with significantly less DNA fragmentation than the use of Botu-Crio® extender at 6 h of incubation, and than the use of either Botu-Crio® or Gent® extender at 24 h of incubation (p<0.05). Conclusions: These results suggest that single layer colloidal centrifugation performed with extended or raw semen prior to cryopreservation reduces DNA fragmentation during the first four hours after thawing. Further studies are needed to determine the influence of freezing extenders on equine sperm DNA fragmentation dynamics.
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Publication Date: 2012-12-05 PubMed ID: 23217215PubMed Central: PMC3599590DOI: 10.1186/1751-0147-54-72Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 article presents a study that investigates the effect of different pre-freezing processes and extenders on post-thawed horse sperm DNA integrity. The results suggest that using specific centrifugation protocols before freezing can effectively reduce DNA damage in the thawed sperm.

Objective of the Study

  • The study essentially aims to experiment and compare how two single layer colloidal centrifugation methods—and the use of three commercial freezing extenders—affect the post-thaw chromatic stability of equine sperm samples at different incubation times. In this context, the term ‘chromatin integrity’ refers to the intactness of sperm DNA.

Significance of Centrifugation Protocols and Extenders

  • A significant challenge is the lack of a standardized method for semen cryopreservation, given the varying quality of cryopreservation between equine males.
  • To counter this, different extenders and processing techniques like colloidal centrifugation are utilized to optimize the quality of sperm after thawing.
  • The choice of extenders and pre-cryopreservation centrifugation methods can significantly affect the resultant DNA fragmentation level, thus affecting sperm quality.

Findings from the Research

  • It was found that the levels of DNA fragmentation in sperm samples that had gone through either of the colloidal centrifugation methods were notably lower both immediately after thawing and after incubation for 4 hours at 37°C. This applied when compared to samples that had undergone standard centrifugation.
  • Among the extenders, using InraFreeze® was associated with lesser DNA fragmentation compared to the use of Botu-Crio® at 6 hours of incubation.
  • Additionally, InraFreeze® led to less DNA fragmentation compared to either Botu-Crio® or Gent® at 24 hours incubation.

Conclusions and Further Research

  • The research revealed that pre-cryopreservation single layer colloidal centrifugation, performed either on extended or raw semen, reduces DNA fragmentation during the initial four hours after thawing.
  • However, the authors call for more studies to fully understand how different freezing extenders influence the fragmentation dynamics of equine sperm DNA.

Cite This Article

APA
Gutiérrez-Cepeda L, Fernández A, Crespo F, Ramírez MÁ, Gosálvez J, Serres C. (2012). The effect of two pre-cryopreservation single layer colloidal centrifugation protocols in combination with different freezing extenders on the fragmentation dynamics of thawed equine sperm DNA. Acta Vet Scand, 54(1), 72. https://doi.org/10.1186/1751-0147-54-72

Publication

Acta veterinaria Scandinavica
ISSN: 1751-0147
NlmUniqueID: 0370400
Country: England
Language: English
Volume: 54
Issue: 1
Pages: 72

Researcher Affiliations

Gutiérrez-Cepeda, Luna
  • Animal Medicine and Surgery Deparment, Veterinary Faculty, UCM, Avda, Puerta de Hierro s/n, Ciudad Universitaria, 28040, Madrid, Spain. luna.gutierrez@vet.ucm.es
Fernández, Alvaro
    Crespo, Francisco
      Ramírez, Miguel Ángel
        Gosálvez, Jaime
          Serres, Consuelo

            MeSH Terms

            • Animals
            • Centrifugation / methods
            • Centrifugation / veterinary
            • Chromatin / physiology
            • Colloids
            • Cryopreservation / veterinary
            • Cryoprotective Agents / pharmacology
            • DNA Fragmentation
            • Freezing
            • Horses / physiology
            • Male
            • Sperm Motility
            • Spermatozoa / physiology
            • Time Factors

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            Citations

            This article has been cited 3 times.
            1. Gutiérrez-Cepeda L, Crespo F, Blazquez JC, Serres C. Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation. Animals (Basel) 2023 Jan 22;13(3).
              doi: 10.3390/ani13030382pubmed: 36766271google scholar: lookup
            2. Ortiz I, Dorado J, Morrell J, Gosálvez J, Crespo F, Jiménez JM, Hidalgo M. New approach to assess sperm DNA fragmentation dynamics: Fine-tuning mathematical models. J Anim Sci Biotechnol 2017;8:23.
              doi: 10.1186/s40104-017-0155-7pubmed: 28286648google scholar: lookup
            3. Strassner FM, Demattio L, Siuda M, Malama E, Muffels G, Bollwein H. Relationships Between Metabolism of Cryopreserved Equine Sperm Determined by the Seahorse Analyzer and Sperm Characteristics Measured by Flow Cytometry and Computer-Assisted Analysis of Motility. Vet Sci 2025 Nov 21;12(12).
              doi: 10.3390/vetsci12121109pubmed: 41472089google scholar: lookup
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