Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm
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<sup>®</sup> (Minitub GmbH, Tie...
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MDPI AG
2023-03-01
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| Online Access: | https://www.mdpi.com/2076-2615/13/7/1200 |
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| author | Anežka Málková Filipp Georgijevič Savvulidi Martin Ptáček Karolína Machová Martina Janošíková Szabolcs Nagy Luděk Stádník |
| author_facet | Anežka Málková Filipp Georgijevič Savvulidi Martin Ptáček Karolína Machová Martina Janošíková Szabolcs Nagy Luděk Stádník |
| author_sort | Anežka Málková |
| collection | DOAJ |
| description | 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<sup>®</sup> (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<sup>®</sup> 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. |
| first_indexed | 2024-03-11T05:44:18Z |
| format | Article |
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| language | English |
| last_indexed | 2024-03-11T05:44:18Z |
| publishDate | 2023-03-01 |
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| record_format | Article |
| series | Animals |
| spelling | doaj.art-ec24b5595bac40ea83d84516c858b47b2023-11-17T16:14:10ZengMDPI AGAnimals2076-26152023-03-01137120010.3390/ani13071200Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram SpermAnežka Málková0Filipp Georgijevič Savvulidi1Martin Ptáček2Karolína Machová3Martina Janošíková4Szabolcs Nagy5Luděk Stádník6Department of Animal Science, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, 165 00 Prague, Czech RepublicDepartment of Animal Science, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, 165 00 Prague, Czech RepublicDepartment of Animal Science, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, 165 00 Prague, Czech RepublicDepartment of Genetics and Breeding, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, 165 00 Prague, Czech RepublicDepartment of Animal Science, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, 165 00 Prague, Czech RepublicInstitute of Animal Sciences, Hungarian University of Agriculture and Life Sciences, Georgikon Campus, H8360 Keszthely, HungaryDepartment of Animal Science, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, 165 00 Prague, Czech RepublicThis 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<sup>®</sup> (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<sup>®</sup> 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.https://www.mdpi.com/2076-2615/13/7/1200cryopreservation capacitysperm motilityplasma membrane and acrosome intactnessmitochondrial membrane potential |
| spellingShingle | Anežka Málková Filipp Georgijevič Savvulidi Martin Ptáček Karolína Machová Martina Janošíková Szabolcs Nagy Luděk Stádník Glycerol-Free Equilibration with the Addition of Glycerol Shortly before the Freezing Procedure: A Perspective Strategy for Cryopreservation of Wallachian Ram Sperm Animals cryopreservation capacity sperm motility plasma membrane and acrosome intactness mitochondrial membrane potential |
| 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 | cryopreservation capacity sperm motility plasma membrane and acrosome intactness mitochondrial membrane potential |
| url | https://www.mdpi.com/2076-2615/13/7/1200 |
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