New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification

Equine oocyte vitrification would benefit the growing in vitro embryo production programs, but further optimization of the protocol is necessary to reach clinical efficiency. Therefore, we aimed to perform a direct comparison of non-permeating and permeating cryoprotective agents (CPAs) during the v...

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Main Authors: Daniel Angel-Velez, Tine De Coster, Nima Azari-Dolatabad, Andrea Fernandez-Montoro, Camilla Benedetti, Osvaldo Bogado Pascottini, Henri Woelders, Ann Van Soom, Katrien Smits
Format: Article
Language:English
Published: MDPI AG 2021-10-01
Series:Animals
Subjects:
Online Access:https://www.mdpi.com/2076-2615/11/11/3077
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author Daniel Angel-Velez
Tine De Coster
Nima Azari-Dolatabad
Andrea Fernandez-Montoro
Camilla Benedetti
Osvaldo Bogado Pascottini
Henri Woelders
Ann Van Soom
Katrien Smits
author_facet Daniel Angel-Velez
Tine De Coster
Nima Azari-Dolatabad
Andrea Fernandez-Montoro
Camilla Benedetti
Osvaldo Bogado Pascottini
Henri Woelders
Ann Van Soom
Katrien Smits
author_sort Daniel Angel-Velez
collection DOAJ
description Equine oocyte vitrification would benefit the growing in vitro embryo production programs, but further optimization of the protocol is necessary to reach clinical efficiency. Therefore, we aimed to perform a direct comparison of non-permeating and permeating cryoprotective agents (CPAs) during the vitrification and warming of equine immature oocytes. In the first experiment, cumulus oocytes complexes (COCs) were vitrified comparing sucrose, trehalose, and galactose in combination with ethylene glycol (EG) and dimethyl sulfoxide (DMSO). In the second experiment, the COCs were vitrified using three mixtures of permeating CPAs in a 50:50 volume ratio (ethylene glycol-dimethyl sulfoxide (ED), propylene glycol-ethylene glycol (PE), and propylene glycol-dimethyl sulfoxide (PD)) with galactose and warmed in different galactose concentrations (0.3 or 0.5 mol/L). Overall, all the treatments supported blastocyst formation, but the developmental rates were lower for all the vitrified groups in the first (4.3 to 7.6%) and the second (3.5 to 9.4%) experiment compared to the control (26.5 and 34.2%, respectively; <i>p</i> < 0.01). In the first experiment, the maturation was not affected by vitrification. The sucrose exhibited lower cleavage than the control (<i>p</i> = 0.02). Although the galactose tended to have lower maturation than trehalose (<i>p</i> = 0.060) and control (<i>p</i> = 0.069), the highest numerical cleavage and blastocyst rates were obtained with this CPA. In the second experiment, the maturation, cleavage, and blastocyst rates were similar between the treatments. Compared to the control, only the ED reached similar maturation (<i>p</i> = 0.02) and PE similar cleavage (<i>p</i> = 0.1). The galactose concentration during warming did not affect the maturation, cleavage, or blastocyst rates (<i>p</i> > 0.1), but the PE-0.3 exhibited the highest blastocyst rate (15.1%) among the treatments, being the only one comparable to the control (34.2%). As such, PE–galactose provides a valuable option for equine immature oocyte vitrification and should be considered for the future optimization of the protocol.
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spelling doaj.art-d1a2155f14ce437fa5f3fd91f5c4915f2023-12-03T13:27:15ZengMDPI AGAnimals2076-26152021-10-011111307710.3390/ani11113077New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte VitrificationDaniel Angel-Velez0Tine De Coster1Nima Azari-Dolatabad2Andrea Fernandez-Montoro3Camilla Benedetti4Osvaldo Bogado Pascottini5Henri Woelders6Ann Van Soom7Katrien Smits8Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartment of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartment of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartment of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartment of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartment of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumWageningen Livestock Research, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The NetherlandsDepartment of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartment of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumEquine oocyte vitrification would benefit the growing in vitro embryo production programs, but further optimization of the protocol is necessary to reach clinical efficiency. Therefore, we aimed to perform a direct comparison of non-permeating and permeating cryoprotective agents (CPAs) during the vitrification and warming of equine immature oocytes. In the first experiment, cumulus oocytes complexes (COCs) were vitrified comparing sucrose, trehalose, and galactose in combination with ethylene glycol (EG) and dimethyl sulfoxide (DMSO). In the second experiment, the COCs were vitrified using three mixtures of permeating CPAs in a 50:50 volume ratio (ethylene glycol-dimethyl sulfoxide (ED), propylene glycol-ethylene glycol (PE), and propylene glycol-dimethyl sulfoxide (PD)) with galactose and warmed in different galactose concentrations (0.3 or 0.5 mol/L). Overall, all the treatments supported blastocyst formation, but the developmental rates were lower for all the vitrified groups in the first (4.3 to 7.6%) and the second (3.5 to 9.4%) experiment compared to the control (26.5 and 34.2%, respectively; <i>p</i> < 0.01). In the first experiment, the maturation was not affected by vitrification. The sucrose exhibited lower cleavage than the control (<i>p</i> = 0.02). Although the galactose tended to have lower maturation than trehalose (<i>p</i> = 0.060) and control (<i>p</i> = 0.069), the highest numerical cleavage and blastocyst rates were obtained with this CPA. In the second experiment, the maturation, cleavage, and blastocyst rates were similar between the treatments. Compared to the control, only the ED reached similar maturation (<i>p</i> = 0.02) and PE similar cleavage (<i>p</i> = 0.1). The galactose concentration during warming did not affect the maturation, cleavage, or blastocyst rates (<i>p</i> > 0.1), but the PE-0.3 exhibited the highest blastocyst rate (15.1%) among the treatments, being the only one comparable to the control (34.2%). As such, PE–galactose provides a valuable option for equine immature oocyte vitrification and should be considered for the future optimization of the protocol.https://www.mdpi.com/2076-2615/11/11/3077cryoprotective agentsequineICSIoocytevitrificationwarming
spellingShingle Daniel Angel-Velez
Tine De Coster
Nima Azari-Dolatabad
Andrea Fernandez-Montoro
Camilla Benedetti
Osvaldo Bogado Pascottini
Henri Woelders
Ann Van Soom
Katrien Smits
New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification
Animals
cryoprotective agents
equine
ICSI
oocyte
vitrification
warming
title New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification
title_full New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification
title_fullStr New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification
title_full_unstemmed New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification
title_short New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification
title_sort new alternative mixtures of cryoprotectants for equine immature oocyte vitrification
topic cryoprotective agents
equine
ICSI
oocyte
vitrification
warming
url https://www.mdpi.com/2076-2615/11/11/3077
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