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Theriogenology2022; 188; 116-124; doi: 10.1016/j.theriogenology.2022.05.022

Extracellular vesicles from follicular fluid may improve the nuclear maturation rate of in vitro matured mare oocytes.

Abstract: The in vitro maturation (IVM) of equine oocytes is still not efficient and does not yield consistent results. The specific requirements of equine oocytes during this process are still largely unknown, which hinders the development of assisted reproductive techniques (ART) in this species. Because the ovarian follicle microenvironment supports oocytes in their acquisition of developmental competence, follicular fluid seems to be a substantial source of bioactive factors that could support the IVM process. Extracellular vesicles (EVs) are cell-secreted molecules in body fluids that are able to deliver molecular signals and transfer genetic information (mRNA, miRNA) between donor and recipient cells. Hence, our hypothesis is that follicular fluid EVs (ffEVs) from small (<20 mm) ovarian follicles can improve the in vitro maturation rate of mare oocytes. To test our hypothesis, equine ovarian follicular fluid was aspirated and ffEVs were isolated by ultracentrifugation, then characterized using nanoparticle tracking analysis and flow cytometry. Additionally, ffEVs were labeled using the ExoGlow-protein EV labeling kit (System Biosciences, Palo Alto, CA). Cumulus-oocyte complexes (COCs) were matured using a one-step method (Method I, continuous culture for 24-38 h) or a two-step method (Method II, initial denudation after 24 h), in the presence (200 μg protein/ml) or absence of ffEVs. The results show the internalization of ffEVs by equine cumulus cells and, for the first time, also by oocytes. The ffEV treatment during two-step culture had a positive effect on the maturation rate of compacted COCs compared to the control group (45.7% and 20.5%, respectively; p < 0.05). No effect of supplementation was observed on the maturation rate during one-step culture. Our results indicate that the supplementation of culture media with EVs isolated from the follicular fluid of small follicles can improve the IVM rate of mare oocytes, suggesting that ffEVs play an important role during this process and may enhance the development of equine ART.
Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.
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Publication Date: 2022-05-27 PubMed ID: 35689941DOI: 10.1016/j.theriogenology.2022.05.022Google 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 explores how extracellular vesicles (EVs) from follicular fluid can potentially enhance the in vitro maturation (IVM) rate of horse (mare) eggs or oocytes, which could result in further improvement of assisted reproductive techniques (ART) in horses.

Introduction and Hypothesis

  • The study was embarked upon in response to inconsistencies observed in the in vitro maturation of equine oocytes. These inconsistencies pose a hindrance to advancements in assisted reproductive techniques.
  • The researchers hypothesized that follicular fluid EVs (ffEVs) drawn from small ovarian follicles (less than 20mm in size) could potentially improve the in vitro maturation rate of mare oocytes.

Methodology

  • Equine ovarian follicular fluid was aspirated, and the ffEVs were isolated by performing ultracentrifugation. The isolated EVs were characterized using nanoparticle tracking analysis and flow cytometry.
  • The researchers also labeled ffEVs using an ExoGlow-protein EV labeling kit. Cumulus-oocyte complexes (COCs) were matured using two methods: a one-step method that entails continuous culture for 24-38 hours, and a two-step method with initial denudation after 24 hours.
  • These maturing processes were done either in the presence of ffEVs (200 μg protein/ml) or without.

Results and Importance

  • The findings indicated that ffEVs were ingested by equine cumulus cells and surprisingly by oocytes for the first time. They observed that the ffEV treatment during the two-step culture positively affected the maturation rate of compacted COCs as opposed to the control group
  • However, there was no observed effect of the ffEV supplementation on the maturation rate during the one-step culture.
  • While the research presents significant results, it also indicates areas requiring further study, such as the influence of follicular size on the presence and concentration of follicular fluid extracellular vesicles, and the specific role these vesicles play in the maturation of oocytes.
  • The results demonstrate the potential of ffEVs supplementation to improve the in vitro maturation rate of mare oocytes. These could be substantial findings for both equine assisted reproduction and the broader field of veterinary science.

Cite This Article

APA
Gabryś J, Kij-Mitka B, Sawicki S, Kochan J, Nowak A, Łojko J, Karnas E, Bugno-Poniewierska M. (2022). Extracellular vesicles from follicular fluid may improve the nuclear maturation rate of in vitro matured mare oocytes. Theriogenology, 188, 116-124. https://doi.org/10.1016/j.theriogenology.2022.05.022

Publication

Theriogenology
ISSN: 1879-3231
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 188
Pages: 116-124

Researcher Affiliations

Gabryś, Julia
  • University of Agriculture in Krakow, Department of Animal Reproduction, Anatomy and Genomics, Mickiewicza 24/28, 30-059, Krakow, Poland. Electronic address: juliagabrys@gmail.com.
Kij-Mitka, Barbara
  • University of Agriculture in Krakow, Department of Animal Reproduction, Anatomy and Genomics, Mickiewicza 24/28, 30-059, Krakow, Poland.
Sawicki, Sebastian
  • University of Agriculture in Krakow, Department of Animal Reproduction, Anatomy and Genomics, Mickiewicza 24/28, 30-059, Krakow, Poland.
Kochan, Joanna
  • University of Agriculture in Krakow, Department of Animal Reproduction, Anatomy and Genomics, Mickiewicza 24/28, 30-059, Krakow, Poland.
Nowak, Agnieszka
  • University of Agriculture in Krakow, Department of Animal Reproduction, Anatomy and Genomics, Mickiewicza 24/28, 30-059, Krakow, Poland.
Łojko, Julianna
  • University of Agriculture in Krakow, Department of Animal Reproduction, Anatomy and Genomics, Mickiewicza 24/28, 30-059, Krakow, Poland.
Karnas, Elżbieta
  • Jagiellonian University, Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Gronostajowa 7, 30-387, Krakow, Poland.
Bugno-Poniewierska, Monika
  • University of Agriculture in Krakow, Department of Animal Reproduction, Anatomy and Genomics, Mickiewicza 24/28, 30-059, Krakow, Poland.

MeSH Terms

  • Animals
  • Cumulus Cells
  • Extracellular Vesicles
  • Female
  • Follicular Fluid
  • Horses
  • In Vitro Oocyte Maturation Techniques / methods
  • In Vitro Oocyte Maturation Techniques / veterinary
  • Oocytes
  • Ovarian Follicle

Conflict of Interest Statement

Declaration of competing interest The authors declare no conflict of interest.

Citations

This article has been cited 23 times.
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