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Home / Equine Research Bank / Equine Vet J / Successful vitrification of equine embryos >300 microns w...
Equine veterinary journal2024; doi: 10.1111/evj.14081

Successful vitrification of equine embryos >300 microns without puncture or aspiration.

Abstract: Equine embryos >300 μm require puncture before vitrification. Protocols that do not require pre-puncture would make vitrification easier and allow for its widespread use. Objective: To design a successful vitrification protocol for embryos >300 μm without puncture as a pre-treatment. Methods: Experimental in vivo study. Methods: Thirty-eight embryos were divided into 3 groups (G1: ≤300 μm, n = 11; G2: >300-500 μm, n = 20; G3: >500 μm, n = 7). Embryos were vitrified using a human vitrification kit. Following a 15 min exposure to equilibration solution (ES; 7.5% DMSO +7.5% ethylene glycol [EG] in a base medium [BM] of M199 HEPES-buffered medium [H199] + hydroxypropyl cellulose + gentamycin), embryos were exposed for ≤90 s to a vitrification solution (15% DMSO +15% EG + 0.5 M trelahose in BM), loaded onto a Cryolock and plunged into LN2. Warming was undertaken by plunging the Cryolock tip into 1 mL of H199 + 20% FBS + pen/strep +1 M sucrose at 42°C for 1 min. The embryos were then moved to a 0.5 M sucrose solution for 4 min, then placed in Vigro Hold for 4 min prior to transfer to a recipient. Results: Pregnancy rates were 81.8% (9/11) for G1, 80% (16/20) for G2, and 0% (0/7) for G3. The largest embryo to survive was 480 μm. Conclusions: Limited numbers and only one pregnancy was followed to term. Conclusions: Equine embryos ≤480 μm can be successfully vitrified using a protocol with a longer exposure time to the ES. This does not appear to have a negative effect on early embryonic development.
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Publication Date: 2024-03-07 PubMed ID: 38450769DOI: 10.1111/evj.14081Google 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 aims to create a successful vitrification process for horse embryos that are greater than 300 micrometers in size, without having to puncture them beforehand, thus making the process simpler to implement.

Research Overview

  • The study dealt with testing a novel vitrification process on horse embryos. Vitrification is a method of freezing embryos to retain their viability and it requires precise control over the process to prevent damage. Usually, horse embryos greater than 300 micrometers in size require a puncture before they can be vitrified. The objective of this study, however, was to find an efficient vitrification process that didn’t involve puncturing the embryo as a pre-treatment.
  • Three groups of different-sized embryos were used in the study, with the aim of testing the effectiveness of a new vitrification protocol. Thirty-eight embryos were divided into groups according to their sizes; less than or equal to 300 micrometers, between 300 and 500 micrometers, and greater than 500 micrometers.

    • Methods

    • The embryos were vitrified using a kit designed for human embryo vitrification. However, the process was altered to fit the requirements of this study.
    • They were initially exposed to an equilibration solution (ES), followed by a vitrification solution. The exposure times for these solutions were meticulously managed to ensure the process didn’t harm the embryos. Afterwards, these embryos were frozen using liquid nitrogen and warmed up later using a specified procedure.
    • After being warmed, the embryos underwent further treatment and were then transferred to a recipient.

    Results

    • The results displayed varying rates of success. The pregnancy rates for embryos ≤300 μm and those >300 to 500 μm were similar, at roughly 80%. However, there were no successful impregnations from the largest embryo group that are >500 μm.
    • The largest embryo that successfully survived the vitrification process was 480 micrometers.
    • The researchers concluded that the method allows successful vitrification of horse embryos smaller than or equal to 480 micrometers. This alteration to the vitrification process did not appear to harm the early development of the embryos.

Conclusions

  • While the sample size was limited and only one pregnancy was followed all the way to term, the study was able to achieve its main objective.
  • Further research might look into refining the approach for larger embryos or examining why the treatment was unsuccessful for the largest group in this study.

Cite This Article

APA
Kovacsy S, Ismer A, Funes J, Hoogewijs M, Wilsher S. (2024). Successful vitrification of equine embryos >300 microns without puncture or aspiration. Equine Vet J. https://doi.org/10.1111/evj.14081

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English

Researcher Affiliations

Kovacsy, Sofia
  • Sharjah Equine Hospital, Sharjah, UAE.
Ismer, Ann
  • Sharjah Equine Hospital, Sharjah, UAE.
Funes, Javier
  • Sharjah Equine Hospital, Sharjah, UAE.
Hoogewijs, Maarten
  • Sharjah Equine Hospital, Sharjah, UAE.
Wilsher, Sandra
  • Sharjah Equine Hospital, Sharjah, UAE.

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Citations

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