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Home / Equine Research Bank / Reprod Domest Anim / Effect of permeable cryoprotectant-free vitrification on DNA...
Reproduction in domestic animals = Zuchthygiene2019; 54 Suppl 3; 53-56; doi: 10.1111/rda.13491

Effect of permeable cryoprotectant-free vitrification on DNA fragmentation of equine oocyte-cumulus cells.

Abstract: DNA fragmentation of cumulus cells could be used as an indicator of oocyte vitrification success as an indirect indicator of the quality of the oocyte. This study was designed to compare the DNA fragmentation of post-mortem equine cumulus cells before or after vitrification in the absence of permeable cryoprotectant agents. Cumulus-oocyte complexes (COCs; n = 56) were recovered from slaughterhouse ovaries and subjected to in vitro maturation (42 hr/38.2°C/5%CO ) before (control group) or after a permeable cryoprotectant-free vitrification method using 1 M sucrose (vitrification group). After in vitro maturation, COCs were denuded, and cumulus cells were washed and stored at -80°C until thawing. Cumulus cell samples were processed with the chromatin dispersion test (Ovoselect, Halotech DNA, Spain). Low, high and total DNA fragmentation percentages of cumulus cells were recorded and compared between the two groups by Student's t test. Results were expressed as mean ± SEM. The vitrified group resulted in significantly higher (p < 0.05) percentages for low (16.81 ± 1.62 vs. 6.63 ± 0.77) and total (21.14 ± 1.84 vs. 12.76 ± 1.48) DNA fragmentation of cumulus cells. There were no significant differences between groups for high DNA fragmentation of cumulus cells. In conclusion, permeable cryoprotectant-free vitrification of equine oocytes increased the total DNA fragmentation rate of cumulus cells but protected them against high DNA fragmentation rates. Further studies are needed to examine the relationship between DNA fragmentation of cumulus cells and the developmental competence of equine oocytes.
© 2019 Blackwell Verlag GmbH.
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Publication Date: 2019-09-13 PubMed ID: 31512317DOI: 10.1111/rda.13491Google Scholar: Lookup
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  • Journal Article

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.

The research paper investigates how a specific type of oocyte preservation method, known as cryoprotectant-free vitrification, affects the DNA fragmentation in equine cumulus cells.

Research Methodology

  • The study involves testing of cumulus-oocyte complexes (COCs) recovered from ovary tissues obtained from a slaughterhouse.
  • The collected COCs were subjected to a maturation process in vitro, conducted at specified conditions (42 hours at 38.2°C with 5% CO2).
  • Two sets of COCs – one from the control group and another from the vitrification group – were selected. The latter underwent a process of permeable cryoprotectant-free vitrification using 1M sucrose.
  • Post-maturation, the COCs were stripped of cells, washed and preserved at -80 degrees Celsius until subsequent thawing.
  • The method used for assessing DNA fragmentation in cumulus cells was the chromatin dispersion test (Ovoselect by Halotech DNA).

Research Findings

  • The study found that the vitrification group displayed a significantly higher rate of low and total DNA fragmentation in the cumulus cells when compared to the control group.
  • In terms of high DNA fragmentation, there were no noticeable differences between the groups.
  • Therefore, the study concluded that the permeable cryoprotectant-free vitrification of equine oocytes led to an increase in the overall DNA fragmentation rate of cumulus cells. However, it protected against high DNA fragmentation rates.

Research Implications and Future Work

  • This research shows the potential of using DNA fragmentation in cumulus cells as an indirect measure of the success of oocyte vitrification.
  • However, the study recommends further research to establish a clear connection between DNA fragmentation in cumulus cells and the developmental competence of equine oocytes.

Cite This Article

APA
Pereira BC, Ortiz I, Dorado JM, Diaz-Jimenez MA, Consuegra C, Gosalvez J, Hidalgo M. (2019). Effect of permeable cryoprotectant-free vitrification on DNA fragmentation of equine oocyte-cumulus cells. Reprod Domest Anim, 54 Suppl 3, 53-56. https://doi.org/10.1111/rda.13491

Publication

Reproduction in domestic animals = Zuchthygiene
ISSN: 1439-0531
NlmUniqueID: 9015668
Country: Germany
Language: English
Volume: 54 Suppl 3
Pages: 53-56

Researcher Affiliations

Pereira, Blasa Carmen
  • Veterinary Reproduction Group, Veterinary Faculty, Department of Animal Medicine and Surgery, University of Cordoba, Cordoba, Spain.
Ortiz, Isabel
  • Veterinary Reproduction Group, Veterinary Faculty, Department of Animal Medicine and Surgery, University of Cordoba, Cordoba, Spain.
Dorado, Jesús Manuel
  • Veterinary Reproduction Group, Veterinary Faculty, Department of Animal Medicine and Surgery, University of Cordoba, Cordoba, Spain.
Diaz-Jimenez, Maria Angeles
  • Veterinary Reproduction Group, Veterinary Faculty, Department of Animal Medicine and Surgery, University of Cordoba, Cordoba, Spain.
Consuegra, Cesar
  • Veterinary Reproduction Group, Veterinary Faculty, Department of Animal Medicine and Surgery, University of Cordoba, Cordoba, Spain.
Gosalvez, Jaime
  • Department of Biology, Autonomous University of Madrid, Madrid, Spain.
Hidalgo, Manuel
  • Veterinary Reproduction Group, Veterinary Faculty, Department of Animal Medicine and Surgery, University of Cordoba, Cordoba, Spain.

MeSH Terms

  • Animals
  • Cryopreservation / methods
  • Cryopreservation / veterinary
  • Cumulus Cells
  • DNA Fragmentation
  • Female
  • Horses
  • Oocytes
  • Vitrification

Grant Funding

  • AGL2013-42726-R / MINECO (Ministerio de Economu00eda y Competitividad), Spain
  • ceiA3 (Andalucu00eda, Spain)
  • II Plan Propio Galileo de Innovaciu00f3n y Transferencia (University of Cordoba, Spain)

References

This article includes 15 references
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Citations

This article has been cited 4 times.
  1. Ortiz I, Dorado J, Pereira B, Diaz-Jimenez M, Consuegra C, Gosalvez J, Hidalgo M. DNA fragmentation of equine cumulus cells from Cumulus-Oocyte complexes submitted to vitrification and its relationship to the developmental competence of the oocyte. Reprod Domest Anim 2022 Oct;57 Suppl 5(Suppl 5):64-67.
    doi: 10.1111/rda.14197pubmed: 35770452google scholar: lookup
  2. López A, Betancourt M, Ducolomb Y, Rodríguez JJ, Casas E, Bonilla E, Bahena I, Retana-Márquez S, Juárez-Rojas L, Casillas F. DNA damage in cumulus cells generated after the vitrification of in vitro matured porcine oocytes and its impact on fertilization and embryo development. Porcine Health Manag 2021 Oct 18;7(1):56.
    doi: 10.1186/s40813-021-00235-wpubmed: 34663451google scholar: lookup
  3. Akkoç T. In Vıtro Productıon of Bovıne Blastocysts and Cryopreservatıon Technıques. Adv Exp Med Biol 2025;1499:51-70.
    doi: 10.1007/5584_2025_868pubmed: 40652129google scholar: lookup
  4. Khajedehi N, Fathi R, Akbarinejad V, Gourabi H. Oocyte Vitrification Reduces its Capability to Repair Sperm DNA Fragmentation and Impairs Embryonic Development. Reprod Sci 2024 May;31(5):1256-1267.
    doi: 10.1007/s43032-023-01419-1pubmed: 38151654google scholar: lookup
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