Comparison of different sucrose-based extenders for stallion sperm vitrification in straws.
Abstract: Vitrification of sperm is based on high-speed freezing by direct exposure to liquid nitrogen using non-permeable cryoprotectants, mainly disaccharides; yet, the concentration of cryoprotectants has a species-specific effect on the sperm cell. The aim of this study was to assess different sucrose concentrations for stallion sperm vitrification. Semen samples (n = 9) were collected from three stallions, centrifuged and resuspended to a concentration of 50 × 10 sperm/ml in a base extender (INRA96 + 1% of bovine serum albumin) with three different sucrose concentrations (Molar): 20 mM (S1), 100 mM (S2), or 200 mM (S3). Then, sperm were filled in covered 0.25 ml straws and directly plunged into liquid nitrogen. For warming, 0.25 ml straw was pulled out the covering straw and immersed in 3 ml of INRA96 at 43°C, with gentle pipetting to accelerate the melting. Total (TM, %) and progressive sperm motility (PM, %) were analysed using computer-assisted sperm analysis. Plasma (PMI, %) and acrosome membrane integrity (AIS, %) were assessed under epifluorescence microscopy. Post-warmed sperm parameters were compared between treatments by ANOVA. S2 showed significantly higher values in comparison with S1 and S3 for TM (S2 = 54.7 ± 5.5 ; S1 = 29.1 ± 3.3 ; S3 = 28.6 ± 3.0 ; p < 0.001) and PM (S2 = 31.3 ± 3.8 ; S1 = 18.5 ± 2.6 ; S3 = 17.7 ± 2.9 ; p < 0.01), respectively. No significant differences were found among treatments for PMI (S2 = 70.3 ± 5.2; S1 = 67.4 ± 4.3; S3 = 70.0 ± 3.7) neither for AIS (S2 = 57.1 ± 3.9; S1 = 53.9 ± 4.2; S3 = 57.0 ± 7.9). In conclusion, a concentration of 100 mM sucrose is recommended for stallion sperm vitrification in straws.
© 2018 Blackwell Verlag GmbH.
Publication Date: 2018-09-22 PubMed ID: 30238663DOI: 10.1111/rda.13265Google Scholar: Lookup
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- Comparative Study
- Journal Article
Summary
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This study compares different concentrations of sucrose used as extenders in the vitrification (rapid freezing) process of horse sperm, demonstrating that a concentration of 100 millimolars (mM) of sucrose is most beneficial for retaining sperm function.
Introduction to the Study
- The researchers focused on studying the effect of various concentrations of sucrose on the vitrification of stallion sperm.
- Vitrification is a process that rapidly freezes sperm, preserving it for future use.
- The nature of cryoprotectants used, primarily disaccharides such as sucrose, is species-specific and affects the outcome of sperm cell preservation.
Methods of the Study
- Semen samples were collected from three stallions and diluted with a base extender to obtain a concentration of 50 × 106 sperm/ml.
- Three different concentrations of sucrose were used: 20 mM (S1), 100 mM (S2), and 200 mM (S3).
- These sperm samples were directly plunged into liquid nitrogen to create a rapid freezing effect.
- To warm the sperm, each straw was immersed in a solution at 43°C and gently pipetted to facilitate melting.
- Post-warming, the researchers analysed the total and progressive sperm motility, plasma and acrosome membrane integrity using computer-assisted sperm analysis and epifluorescence microscopy.
Results of the Study
- The sperm vitrified with 100 mM sucrose (S2) showed significantly higher total and progressive motility rates compared to those vitrified with 20 mM (S1) and 200 mM (S3) sucrose.
- No significant difference was found among the three groups with regards to plasma membrane integrity and acrosome membrane integrity.
Conclusion
- Based on their findings, the researchers recommend using a concentration of 100 mM sucrose for the vitrification of stallion sperm.
- This concentration is found to maintain higher total and progressive motility, which indicates the efficiency of 100 mM sucrose as a cryoprotectant during the vitrification process.
Cite This Article
APA
Consuegra C, Crespo F, Dorado J, Ortiz I, Diaz-Jimenez M, Pereira B, Hidalgo M.
(2018).
Comparison of different sucrose-based extenders for stallion sperm vitrification in straws.
Reprod Domest Anim, 53 Suppl 2, 59-61.
https://doi.org/10.1111/rda.13265 Publication
Researcher Affiliations
- Veterinary Reproduction Group, University of Cordoba, Cordoba, Spain.
- Department of Reproduction, Centro Militar de Cría Caballar, Avila, Spain.
- Veterinary Reproduction Group, University of Cordoba, Cordoba, Spain.
- Veterinary Reproduction Group, University of Cordoba, Cordoba, Spain.
- Veterinary Reproduction Group, University of Cordoba, Cordoba, Spain.
- Veterinary Reproduction Group, University of Cordoba, Cordoba, Spain.
- Veterinary Reproduction Group, University of Cordoba, Cordoba, Spain.
MeSH Terms
- Animals
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents / pharmacology
- Horses
- Male
- Semen Analysis / veterinary
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sperm Motility / drug effects
- Sucrose / pharmacology
- Vitrification
Citations
This article has been cited 1 times.- Hidalgo M, Ortiz I. Sperm Vitrification in Horse and Donkey. Methods Mol Biol 2025;2897:137-145.
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