Cryopreservation of stallion spermatozoa using different cryoprotectants and combinations of cryoprotectants.
Abstract: The present study investigates the effects of five cryoprotectants (CPAs) and cryoprotectant combinations on the post-thaw total motility, progressive motility, viability, mitochondrial membrane potential and acrosome integrity in stallion spermatozoa. In Experiment 1, the objective was to compare the impact of different concentrations (2.5%, 3.5% and 5%) of a single CPA, including glycerol (Gly), ethylene glycol (EG), dimethyl sulphoxide (DMSO), methyl formamide (MF), and dimethylformamide (DMF) for stallion spermatozoa cryopreservation. In Experiment 2, two or more CPAs were used to assess whether this improved post-thaw spermatozoa quality. Gly, MF and DMF, were used to prepare seven combinations of freezing extender with different mixtures of cryoprotectant, and the 3.5% Gly, MF and DMF were used as a control group. The results show that post-thaw total motility, progressive motility, viability, and mitochondrial membrane potential for all concentrations of EG and DMSO were less than the 3.5% and 5% Gly and MF and 2.5% and 3.5% DMF (P<0.05). Use of the 3.5% concentration resulted in the greater post-thaw total motility and progressive motility than the 2.5% and 5% concentrations for all CPAs. The results for the use of different combinations of cryoprotectant indicate there are differences in progressive motility and viability. The viability with the use of Gly(2/3)+MF(1/3) was 44.65% and was greater than the Gly(1/3)+MF(1/3)+DMF(1/3) (30.96%), MF(2/3)+DMF(1/3) (35.05%), Gly (32.21%) and MF(33.76%) (P<0.05). The progressive motility with the use of the MF(2/3)+Gly(1/3) combination was 36.0% and was greater than in the DMF (25.0%) and MF(2/3)+DMF(1/3) (22.7%) (P<0.05). These results suggest that using the appropriate cryoprotectant combination instead of a single cryoprotectant can improve horse spermatozoa cryopreservation.
Copyright © 2015 Elsevier B.V. All rights reserved.
Publication Date: 2015-10-03 PubMed ID: 26573763DOI: 10.1016/j.anireprosci.2015.09.020Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research explores the effectiveness of different cryoprotectants, and their combinations, on preserving the quality of stallion sperm after freezing and thawing. The study concludes that using the correct combination of cryoprotectants can improve post-thaw quality of the sperm cells.
Methodology
- The study consists of two experiments. In the first experiment, the effects of different concentrations (2.5%, 3.5%, and 5%) of single cryoprotectants were assessed. These included glycerol (Gly), ethylene glycol (EG), dimethyl sulphoxide (DMSO), methyl formamide (MF), and dimethylformamide (DMF).
- In the second experiment, the researchers combined two or more cryoprotectants to see if this improves the post-thaw quality of sperm. They used Gly, MF, and DMF to prepare seven different combinations of freezing extender with different mixtures of cryoprotectant.
Results
- In general, the results show that post-thaw total motility, progressive motility, viability, and mitochondrial membrane potential for all concentrations of EG and DMSO were lesser than the 3.5% and 5% Gly and MF and 2.5% and 3.5% DMF.
- Among the single cryoprotectants used, 3.5% concentration resulted in better post-thaw total motility and progressive motility than the 2.5% and 5% concentrations.
- The results for the use of different combinations of cryoprotectant indicate there are differences in progressive motility and viability. For instance, the viability using a Gly(2/3)+MF(1/3) combination was 44.65%, significantly higher than other combinations.
- The best progressive motility result was found with the combination of MF(2/3)+Gly(1/3), standing at 36.0%, which was significantly greater than using DMF alone or the MF(2/3)+DMF(1/3) combination.
Conclusion
- This study shows that each cryoprotectant or mixture impacts the post-thaw quality of horse sperm differently. Choosing the correct combinations of cryoprotectants may improve the preservation of sperm, potentially benefiting various applications, from breeding to research.
Cite This Article
APA
Wu Z, Zheng X, Luo Y, Huo F, Dong H, Zhang G, Yu W, Tian F, He L, Chen J.
(2015).
Cryopreservation of stallion spermatozoa using different cryoprotectants and combinations of cryoprotectants.
Anim Reprod Sci, 163, 75-81.
https://doi.org/10.1016/j.anireprosci.2015.09.020 Publication
Researcher Affiliations
- College of Animal Science and Technology, Shihezi University, Shihezi 832000, Xinjiang, China.
- Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
- Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
- Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
- Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
- Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
- The Breeding Center of Yili Horse, Zhaosu 835600, Xingjiang, China.
- College of Animal Science and Technology, Shihezi University, Shihezi 832000, Xinjiang, China.
- College of Animal Science and Technology, Shihezi University, Shihezi 832000, Xinjiang, China.
- College of Animal Science and Technology, Shihezi University, Shihezi 832000, Xinjiang, China; Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China. Electronic address: chenjb126@126.com.
MeSH Terms
- Animals
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents / administration & dosage
- Cryoprotective Agents / pharmacology
- Horses / physiology
- Male
- Semen Preservation / methods
- Semen Preservation / veterinary
- Spermatozoa / physiology
Citations
This article has been cited 5 times.- Akhtar MF, Ma Q, Li Y, Chai W, Zhang Z, Li L, Wang C. Effect of Sperm Cryopreservation in Farm Animals Using Nanotechnology. Animals (Basel) 2022 Sep 2;12(17).
- Bustani GS, Baiee FH. Semen extenders: An evaluative overview of preservative mechanisms of semen and semen extenders. Vet World 2021 May;14(5):1220-1233.
- Hungerford AJ, Harrison N, Bakos HW, Aitken RJ. Development of an improved medium for the preservation of human spermatozoa. J Assist Reprod Genet 2025 Jul;42(7):2167-2180.
- Zhang L, Jiang C, Wang X, Sohail T, Sun Y, Sun X, Wang J, Li Y. Freezing Hu ram semen: influence of different penetrating cryoprotectants and egg yolk level on the post-thaw quality of sperm. Anim Biosci 2024 Sep;37(9):1548-1557.
- Li W, Song P, Liu B. [Study on thermophysical properties and effect of lyoprotectants in freezing human hepatoma Hep-G (2) cells]. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi 2019 Oct 25;36(5):803-809.
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