Liposomes as an alternative to egg yolk in stallion freezing extender.
Abstract: Egg yolk is normally used as a protective agent to freeze semen of equine and other species. However, addition of egg yolk in extenders is not without disadvantages and the demand to find cryoprotective alternatives is strong. The objective of this study was to test the cryoprotective capacities of liposomes composed of egg yolk phospholipids. Two experiments were conducted: 1) the first to determine the optimal composition and concentration of liposomes to preserve post-thaw motility and membrane integrity of spermatozoa; 2) the second to assess in vivo the cryoprotective capacities of these liposomes. In Experiment 2, post-thaw motility and membrane integrity of spermatozoa were also analyzed. Experiment 1 demonstrated that liposomes composed of phospholipids E80 (commercial lecithins from egg yolk composed mainly of phosphatidylcholine and phosphatidylethanolamine) and of Hank's salts-glucose-lactose solution (E80-liposomes) were the most efficient in preserving post-thaw motility. The optimal concentration was 4 % (v/v). In Experiment 2, fertility rate after artificial insemination of semen frozen with E80-liposomes was 55 % (22/40) compared with 68 % (27/40) with the control extender containing egg yolk (EY) (p = 0.23). Post-thaw motility parameters were higher with EY than with E80-liposomes (p < 0.0001). For post-thaw membrane integrity no difference was observed between the two extenders (p = 0.08). Liposomes composed of egg yolk phospholipids appeared to be a promising alternative to replace egg yolk in semen freezing extenders in equine species.
Copyright © 2012 Elsevier Inc. All rights reserved.
Publication Date: 2011-09-14 PubMed ID: 21924469DOI: 10.1016/j.theriogenology.2011.08.001Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research examines the use of liposomes (composed of egg yolk phospholipids) as an alternative to egg yolk in extending the freezing capacity of stallion semen. The study found that these liposomes could be a promising alternative, offering similar levels of protection to spermatozoa during freezing.
Objective of Study
- The main purpose of this study was to explore the cryoprotective capacities of liposomes made up of egg yolk phospholipids. The use of egg yolk as a standard protective agent for freezing stallion semen presents certain disadvantages and thus, the research aimed to provide an alternative.
Methodology of the Study
- The research was conducted in two experiments. The first experiment aimed to determine the optimal composition and concentrations of liposomes that would best preserve the post-thaw motility and membrane integrity of the spermatozoa.
- The second experiment assessed the in vivo cryoprotective capacities of these liposomes, and also analyzed the post-thaw motility and membrane integrity of spermatozoa.
Key Findings from the Experiments
- The results from the first experiment revealed that the liposomes made of phospholipids E80 and a solution of Hank’s salts-glucose-lactose were the most effective in preserving post-thaw motility. The optimal concentration for this purpose was found to be 4%.
- The second experiment showed that the fertility rate after artificial insemination of semen frozen with E80-liposomes was 55%, compared to the 68% fertility rate with the control extender that contained egg yolk. However, this difference was not statistically significant.
- The study also found that post-thaw motility parameters were higher with egg yolk than with E80-liposomes, but for post-thaw membrane integrity, there was no difference observed between the two extenders.
Conclusion of the paper
- Based on the results from the experiments, it was concluded that liposomes composed of egg yolk phospholipids present a promising alternative to egg yolk in semen freezing extenders for equine species. This could help to overcome some of the disadvantages associated with the use of egg yolk in freezing semen.
Cite This Article
APA
Pillet E, Labbe C, Batellier F, Duchamp G, Beaumal V, Anton M, Desherces S, Schmitt E, Magistrini M.
(2011).
Liposomes as an alternative to egg yolk in stallion freezing extender.
Theriogenology, 77(2), 268-279.
https://doi.org/10.1016/j.theriogenology.2011.08.001 Publication
Researcher Affiliations
- INRA, UMR85 Physiologie de la Reproduction et des Comportements, Nouzilly, France.
MeSH Terms
- Animals
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents / chemistry
- Egg Yolk / chemistry
- Female
- Horses
- Insemination, Artificial / veterinary
- Liposomes / chemistry
- Male
- Phospholipids
- Pregnancy
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sperm Motility
Citations
This article has been cited 11 times.- Boni R, Ruggiero R, De Luca F, Serritella ML, Di Palma T, Cecchini Gualandi S. Repeatability of Selected Parameters Related to Stallion Sperm Quality and Cryotolerance. Animals (Basel) 2025 Sep 26;15(19).
- de Almeida RA, da Silva LS, de Lima LGF, Lourencetti FA, Freitas-Dell Aqua C, Ferreira JCP, Oba E. Egg-yolk- and a liposome-based extenders: refrigeration time and effects on ram semen quality. Anim Reprod 2025;22(2):e20240052.
- Suyatno S, Herdis H, Priyatno TP, Pangestu M, Santoso S, Kostaman T, Hudaya MF, Sushadi PS, Lupitasari FBI, Hafid A, Muttaqin Z, Nainggolan IM, Sholikin MM, Sitaresmi PI. A comprehensive meta-analysis on the efficacy of growth factor enriched cryo-agents in enhancing post-thaw quality of mammalian semen. Anim Biosci 2025 Oct;38(10):2094-2110.
- Dcunha R, Mutalik SP, Reji RA, Mutalik S, Kalthur SG, Hegde P, Murari MS, Raghu SV, Banerjee S, Kumar A, Adiga SK, Zhao Y, Kannan N, Kalthur G. Liposome-based Freezing Medium Improves the Outcome of Mouse Prepubertal Testicular Tissue Cryopreservation. Reprod Sci 2024 Nov;31(11):3532-3548.
- Boni R, Ruggiero R, Di Palma T, Ferrara MA, Preziosi G, Cecchini Gualandi S. Stallion Sperm Freezing with Different Extenders: Role of Antioxidant Activity and Nitric Oxide Production. Animals (Basel) 2024 Aug 25;14(17).
- Kameni SL, Dlamini NH, Feugang JM. Exploring the full potential of sperm function with nanotechnology tools. Anim Reprod 2024;21(3):e20240033.
- Deng T, Wang W, Fu Z, Xie Y, Zhou Y, Pu J, Chen K, Yao B, Li X, Yao J. Lipidomics random forest algorithm of seminal plasma is a promising method for enhancing the diagnosis of necrozoospermia. Metabolomics 2024 May 21;20(3):57.
- Ozimic S, Ban-Frangez H, Stimpfel M. Sperm Cryopreservation Today: Approaches, Efficiency, and Pitfalls. Curr Issues Mol Biol 2023 May 29;45(6):4716-4734.
- Pernas S, Fernandez-Novo A, Barrajon-Masa C, Mozas P, Pérez-Villalobos N, Martín-Maldonado B, Oliet A, Astiz S, Pérez-Garnelo SS. Bull Semen Obtained on Beef Farms by Electroejaculation: Sperm Quality in the First Two Hours of Storing with Different Extenders and Holding Temperatures. Animals (Basel) 2023 May 6;13(9).
- Saadeldin IM, Khalil WA, Alharbi MG, Lee SH. The Current Trends in Using Nanoparticles, Liposomes, and Exosomes for Semen Cryopreservation. Animals (Basel) 2020 Dec 3;10(12).
- Luna-Orozco JR, González-Ramos MA, Calderón-Leyva G, Gaytán-Alemán LR, Arellano-Rodríguez F, Ángel-García O, Véliz-Deras FG. Comparison of different diluents based on liposomes and egg yolk for ram semen cooling and cryopreservation. Iran J Vet Res 2019 Spring;20(2):126-130.
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