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Theriogenology2007; 68(5); 804-812; doi: 10.1016/j.theriogenology.2007.06.009

Rapidly cooled horse spermatozoa: loss of viability is due to osmotic imbalance during thawing, not intracellular ice formation.

Abstract: The cellular damage that spermatozoa encounter at rapid rates of cooling has often been attributed to the formation of intracellular ice. However, no direct evidence of intracellular ice has been presented. An alternative mechanism has been proposed by Morris (2006) that cell damage is a result of an osmotic imbalance encountered during thawing. This paper examines whether intracellular ice forms during rapid cooling or if an alternative mechanism is present. Horse spermatozoa were cooled at a range of cooling rates from 0.3 to 3,000 degrees C/min in the presence of a cryoprotectant. The ultrastructure of the samples was examined by Cryo Scanning Electron Microscopy (CryoSEM) and freeze substitution, to determine whether intracellular ice formed and to examine alternative mechanisms of cell injury during rapid cooling. No intracellular ice formation was detected at any cooling rate. Differential scanning Calorimetry (DSC) was employed to examine the amount of ice formed at different rate of cooling. It is concluded that cell damage to horse spermatozoa, at cooling rates of up to 3,000 degrees C/min, is not caused by intracellular ice formation. Spermatozoa that have been cooled at high rates are subjected to an osmotic shock when they are thawed.
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Publication Date: 2007-07-23 PubMed ID: 17645937DOI: 10.1016/j.theriogenology.2007.06.009Google 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 research study investigates the causes of sperm cell damage during the rapid cooling process, finding that intracellular ice does not form in horse sperm cells cooled at rates up to 3,000 degrees C/min, and suggests that cell damage is likely due to osmotic imbalance during the thawing process.

Experiment Design and Methodology

  • The research utilized horse sperm cells and subjected them to cooling rates ranging from 0.3 to 3,000 degrees C/min with the inclusion of a cryoprotectant.
  • Cryo Scanning Electron Microscopy (CryoSEM) and freeze substitution techniques were used to examine the impact on cell ultrastructure after cooling, specifically to verify if intracellular ice was formed.
  • Differential scanning calorimetry (DSC) was also utilized to ascertain the quantity of ice formed at varying cooling rates.

Study Findings

  • Upon examination, it was found that no intracellular ice formation occurred at any level of the cooling rates applied. This contradicted the prevalent belief that rapid cooling caused intracellular ice formation, leading to cell damage.
  • Using DSC, the study further reinforced that the amount of ice formed did not differ significantly across the various cooling rates, minimizing the likelihood of intracellular ice damaging the cells.

Conclusions

  • The findings strongly suggest that the primary cause of sperm cell damage in the rapid cooling process isn’t due to intracellular ice formation as commonly thought, but instead potentially due to osmotic imbalance when cells are thawed.
  • This research therefore provides vital insights into cryopreservation processes, potentially directing future studies to focus on managing osmotic imbalance during the thawing process to reduce cell damage.

Cite This Article

APA
Morris GJ, Faszer K, Green JE, Draper D, Grout BW, Fonseca F. (2007). Rapidly cooled horse spermatozoa: loss of viability is due to osmotic imbalance during thawing, not intracellular ice formation. Theriogenology, 68(5), 804-812. https://doi.org/10.1016/j.theriogenology.2007.06.009

Publication

Theriogenology
ISSN: 0093-691X
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 68
Issue: 5
Pages: 804-812

Researcher Affiliations

Morris, G J
  • Asymptote Ltd, St John's Innovation Centre, Cambridge, UK. jmorris@asymptote.co.uk
Faszer, K
    Green, J E
      Draper, D
        Grout, B W W
          Fonseca, F

            MeSH Terms

            • Animals
            • Calorimetry, Differential Scanning
            • Cell Survival
            • Cryopreservation / methods
            • Horses
            • Ice / adverse effects
            • Male
            • Microscopy, Electron, Scanning
            • Osmotic Pressure
            • Semen Preservation / adverse effects
            • Semen Preservation / methods
            • Semen Preservation / veterinary
            • Sperm Motility
            • Spermatozoa / cytology
            • Spermatozoa / metabolism
            • Temperature
            • Time Factors

            Citations

            This article has been cited 20 times.
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