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Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm

SIMPLE SUMMARY: This study investigated the effect of cryoprotectant glycerol addition in different phases of sperm equilibration on sperm motility and viability parameters of thawed ram spermatozoa. The purpose of this study was to compare two strategies of glycerol addition for sperm cryo-preserva...

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Autores principales: Málková, Anežka, Savvulidi, Filipp Georgijevič, Ptáček, Martin, Machová, Karolína, Janošíková, Martina, Nagy, Szabolcs, Stádník, Luděk
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10093609/
https://www.ncbi.nlm.nih.gov/pubmed/37048456
http://dx.doi.org/10.3390/ani13071200
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author Málková, Anežka
Savvulidi, Filipp Georgijevič
Ptáček, Martin
Machová, Karolína
Janošíková, Martina
Nagy, Szabolcs
Stádník, Luděk
author_facet Málková, Anežka
Savvulidi, Filipp Georgijevič
Ptáček, Martin
Machová, Karolína
Janošíková, Martina
Nagy, Szabolcs
Stádník, Luděk
author_sort Málková, Anežka
collection PubMed
description SIMPLE SUMMARY: This study investigated the effect of cryoprotectant glycerol addition in different phases of sperm equilibration on sperm motility and viability parameters of thawed ram spermatozoa. The purpose of this study was to compare two strategies of glycerol addition for sperm cryo-preservation. The first strategy included the use of a glycerol-free extender for the procedure of glycerol-free equilibration and chilling, with the glycerolation of the extender by 6% glycerol shortly before sperm slow freezing. The second strategy included the use of a freezing extender already glycerolated by 6% glycerol before the equilibration and chilling of sperm and following slow freezing. The obtained results were statistically significant and demonstrated that the glycerol-free equilibration with the addition of glycerol at least 10 min before freezing is the best strategy for cryopreservation of Wallachian ram sperm. This represents a perspective strategy in the cryopreservation process for local sheep breeds and indicates a potential strategy to test for ram cryopreservation in a generalized manner. ABSTRACT: This study investigated the effect of glycerol added in different phases of sperm equilibration on CASA and flow cytometry parameters of thawed ram spermatozoa. Sperm was collected from adult Wallachian rams. The freezing extender was glycerol-free ANDROMED(®) (Minitub GmbH, Tiefenbach, Germany) supplied by 6% exogenous glycerol at different stages of the cryopreservation process. The purpose of this study was to compare two strategies of glycerol addition for sperm cryopreservation. The first strategy included the use of a glycerol-free extender for the procedure of glycerol-free equilibration and chilling, with the glycerolation of the extender by 6% glycerol shortly before sperm slow freezing (GFA). The second strategy included the use of a freezing extender already glycerolated by 6% glycerol before the equilibration and chilling of sperm and following slow freezing (GA). Sperm samples were analyzed after equilibration (but before freezing) and after thawing (at T0, T1 h, and T2 h time points). iSperm(®) mCASA (Aidmics Biotechnology Co., LTD., Taipei, Taiwan) was used for the evaluation of sperm kinematics. Flow cytometry was used to measure sperm viability (plasma membrane/acrosome intactness) and mitochondrial membrane potential. The obtained results significantly demonstrated that the glycerol-free equilibration with the addition of glycerol shortly before freezing is a perspective strategy for cryopreservation of Wallachian ram sperm.
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spelling pubmed-100936092023-04-13 Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm Málková, Anežka Savvulidi, Filipp Georgijevič Ptáček, Martin Machová, Karolína Janošíková, Martina Nagy, Szabolcs Stádník, Luděk Animals (Basel) Communication SIMPLE SUMMARY: This study investigated the effect of cryoprotectant glycerol addition in different phases of sperm equilibration on sperm motility and viability parameters of thawed ram spermatozoa. The purpose of this study was to compare two strategies of glycerol addition for sperm cryo-preservation. The first strategy included the use of a glycerol-free extender for the procedure of glycerol-free equilibration and chilling, with the glycerolation of the extender by 6% glycerol shortly before sperm slow freezing. The second strategy included the use of a freezing extender already glycerolated by 6% glycerol before the equilibration and chilling of sperm and following slow freezing. The obtained results were statistically significant and demonstrated that the glycerol-free equilibration with the addition of glycerol at least 10 min before freezing is the best strategy for cryopreservation of Wallachian ram sperm. This represents a perspective strategy in the cryopreservation process for local sheep breeds and indicates a potential strategy to test for ram cryopreservation in a generalized manner. ABSTRACT: This study investigated the effect of glycerol added in different phases of sperm equilibration on CASA and flow cytometry parameters of thawed ram spermatozoa. Sperm was collected from adult Wallachian rams. The freezing extender was glycerol-free ANDROMED(®) (Minitub GmbH, Tiefenbach, Germany) supplied by 6% exogenous glycerol at different stages of the cryopreservation process. The purpose of this study was to compare two strategies of glycerol addition for sperm cryopreservation. The first strategy included the use of a glycerol-free extender for the procedure of glycerol-free equilibration and chilling, with the glycerolation of the extender by 6% glycerol shortly before sperm slow freezing (GFA). The second strategy included the use of a freezing extender already glycerolated by 6% glycerol before the equilibration and chilling of sperm and following slow freezing (GA). Sperm samples were analyzed after equilibration (but before freezing) and after thawing (at T0, T1 h, and T2 h time points). iSperm(®) mCASA (Aidmics Biotechnology Co., LTD., Taipei, Taiwan) was used for the evaluation of sperm kinematics. Flow cytometry was used to measure sperm viability (plasma membrane/acrosome intactness) and mitochondrial membrane potential. The obtained results significantly demonstrated that the glycerol-free equilibration with the addition of glycerol shortly before freezing is a perspective strategy for cryopreservation of Wallachian ram sperm. MDPI 2023-03-29 /pmc/articles/PMC10093609/ /pubmed/37048456 http://dx.doi.org/10.3390/ani13071200 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Málková, Anežka
Savvulidi, Filipp Georgijevič
Ptáček, Martin
Machová, Karolína
Janošíková, Martina
Nagy, Szabolcs
Stádník, Luděk
Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm
title Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm
title_full Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm
title_fullStr Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm
title_full_unstemmed Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm
title_short Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm
title_sort glycerol-free equilibration with the addition of glycerol shortly before the freezing procedure: a perspective strategy for cryopreservation of wallachian ram sperm
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10093609/
https://www.ncbi.nlm.nih.gov/pubmed/37048456
http://dx.doi.org/10.3390/ani13071200
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