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Animals : an open access journal from MDPI2021; 11(11); 3077; doi: 10.3390/ani11113077

New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification.

Abstract: 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; 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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Publication Date: 2021-10-28 PubMed ID: 34827809PubMed Central: PMC8614364DOI: 10.3390/ani11113077Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article explores novel combinations of cryoprotective agents (CPAs) for the vitrification or freezing of equine immature oocytes, an essential step in in vitro equine embryo production. While all tested mixtures catered for blastocyst generation, those discouraged developmental rates compared to the control group. However, the mixture of propylene glycol-ethylene glycol (PE) and galactose notably equaled the developmental rates of the control, suggesting potential for future protocol enhancement.

Aims and Methodology

  • The main objective of this research was to compare the effectiveness of permeating and non-permeating CPAs in the vitrification and warming of equine immature oocytes. This was a bid to optimize the protocol in light of the steadily increasing in vitro embryo production programs.
  • In their experimental setup, the researchers vitrified cumulus oocyte complexes (COCs) using different additives such as sucrose, trehalose, and galactose (non-permeating CPAs) mixed with permeating CPAs like ethylene glycol (EG) and dimethyl sulfoxide (DMSO).
  • In a second experiment, they attempted vitrifying the COCs with three different mixtures of permeating CPAs (EG-DMSO, propylene glycol-EG (PE), and propylene glycol-DMSO) each with galactose, but warmed them in varying galactose concentrations.

Findings

  • All treatments yielded blastocysts, an early embryonic form signalling successful fertilization. However, the blastocyst developmental rates conveyed by the new combinations was lower than those of the control, or standard, method.
  • In terms of maturation, vitrification (the process of transitioning to an amorphous solid state) did not significantly impact the results in the first experiment, but sucrose led to a lower cleavage (early embryo development) rate than the control. Galactose, on the other hand, while registering lower maturation rates than trehalose and control, saw the maximum numerical cleavage and blastocyst rates.
  • In the second experiment, maturation, cleavage, and blastocyst rates remained consistent across treatments. Of the possible pairings, only EG-DMSO equaled the control’s maturation rates, and PE equalled the controls cleavage.
  • The concentration of galactose during warming had no effect on the rates of maturation, cleavage, or blastocyst formation. Yet, the PE-galactose mixture at a 0.3 mol/L concentration showed the highest blastocyst rate of all treatments, comparable to the control.

Conclusions

  • The research suggests that the PE-galactose mixture, particularly at a concentration of 0.3 mol/L, provides a promising option for vitrification of equine immature oocytes and merits further examination in the quest to refine the protocol.

Cite This Article

APA
Angel-Velez D, De Coster T, Azari-Dolatabad N, Fernandez-Montoro A, Benedetti C, Bogado Pascottini O, Woelders H, Van Soom A, Smits K. (2021). New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification. Animals (Basel), 11(11), 3077. https://doi.org/10.3390/ani11113077

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 11
Issue: 11
PII: 3077

Researcher Affiliations

Angel-Velez, Daniel
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
  • Research Group in Animal Sciences-INCA-CES, Universidad CES, Medellin 050021, Colombia.
De Coster, Tine
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Azari-Dolatabad, Nima
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Fernandez-Montoro, Andrea
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Benedetti, Camilla
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Bogado Pascottini, Osvaldo
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
  • Veterinary Physiology and Biochemistry, Department of Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Belgium.
Woelders, Henri
  • Wageningen Livestock Research, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
Van Soom, Ann
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Smits, Katrien
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

This article has been cited 6 times.
  1. Angel-Velez D, Meese T, Hedia M, Fernandez-Montoro A, De Coster T, Pascottini OB, Van Nieuwerburgh F, Govaere J, Van Soom A, Pavani K, Smits K. Transcriptomics Reveal Molecular Differences in Equine Oocytes Vitrified before and after In Vitro Maturation. Int J Mol Sci 2023 Apr 7;24(8).
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