An improved vitrification protocol for equine immature oocytes, resulting in a first live foal.
Abstract: The success rate for vitrification of immature equine oocytes is low. Although vitrified-warmed oocytes are able to mature, further embryonic development appears to be compromised. Objective: The aim of this study was to compare two vitrification protocols, and to examine the effect of the number of layers of cumulus cells surrounding the oocyte during vitrification of immature equine oocytes. Methods: Experimental in vitro and in vivo trials. Methods: Immature equine oocytes were vitrified after a short exposure to high concentrations of cryoprotective agents (CPAs), or a long exposure to lower concentrations of CPAs. In Experiment 1, the maturation of oocytes surrounded by multiple layers of cumulus cells (CC oocytes) and oocytes surrounded by only corona radiata (CR oocytes) was investigated. In Experiment 2, spindle configuration was determined for CR oocytes vitrified using the two vitrification protocols. In Experiment 3, further embryonic development was studied after fertilisation and culture. Embryo transfer was performed in a standard manner. Results: Similar nuclear maturation rates were observed for CR oocytes vitrified using the long exposure and nonvitrified controls. Furthermore, a lower maturation rate was obtained for CC oocytes vitrified with the short exposure compared to control CR oocytes (P = 0.001). Both vitrification protocols resulted in significantly higher rates of aberrant spindle configuration than the control groups (P<0.05). Blastocyst development only occurred in CR oocytes vitrified using the short vitrification protocol, and even though blastocyst rates were significantly lower than in the control group (P<0.001), transfer of five embryos resulted in one healthy foal. Conclusions: The relatively low number of equine oocytes and embryo transfer procedures performed. Conclusions: For vitrification of immature equine oocytes, the use of 1) CR oocytes, 2) a high concentration of CPAs, and 3) a short exposure time may be key factors for maintaining developmental competence.
© 2017 EVJ Ltd.
Publication Date: 2017-09-21 PubMed ID: 28833413DOI: 10.1111/evj.12747Google Scholar: Lookup
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- Journal Article
Summary
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The study was focused on improving the efficiency of handling immature equine oocytes through vitrification, with the ultimate aim of successful embryonic development. It involved comparing different vitrification protocols and assessing the impact of varying the number of cumulus cells surrounding the oocyte during vitrification.
Methods
The research employed both in vitro and in vivo trials, utilizing immature equine oocytes. There were two specific vitrification protocols compared:
- Short exposure to high concentrations of cryoprotective agents (CPAs)
- Long exposure to lower concentrations of CPAs
Three experiments were conducted. The first one inspected the maturation of oocytes surrounded by multiple layers of cumulus cells and oocytes surrounded by corona radiata only. The second experiment determined spindle configuration for oocytes surrounded by corona radiata using both vitrification protocols. The final experiment observed further embryonic development after fertilisation and culture, with standard embryo transfer procedures adhered to.
Results
The study found similar nuclear maturation rates for oocytes surrounded by corona radiata when subjected to long exposure to lower concentrations of CPAs and non-vitrified controls. However, oocytes surrounded by multiple layers of cumulus cells exposed to short exposure to high concentrations of CPAs exhibited lower maturation rates than control corona radiata oocytes. Additionally, higher rates of aberrant spindle configuration were identified in both vitrification protocols compared to controls. Nonetheless, blastocyst development only occurred in corona radiata oocytes vitrified using the short exposure high concentration CPA protocol. Although the blastocyst rates were lower than in the control groups, the transfer of five embryos resulted in one healthy foal.
Conclusion
The research concluded that for optimal vitrification of immature equine oocytes, it is essential to embed key factors including:
- Using corona radiata oocytes
- Employing a high concentration of Cryoprotective agents
- Ensuring a short exposure time
These may enhance the maintenance of developmental competence even though the study noted a relatively low number of equine oocytes and embryo transfer procedures.
Cite This Article
APA
Ortiz-Escribano N, Bogado Pascottini O, Woelders H, Vandenberghe L, De Schauwer C, Govaere J, Van den Abbeel E, Vullers T, Ververs C, Roels K, Van De Velde M, Van Soom A, Smits K.
(2017).
An improved vitrification protocol for equine immature oocytes, resulting in a first live foal.
Equine Vet J, 50(3), 391-397.
https://doi.org/10.1111/evj.12747 Publication
Researcher Affiliations
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
- Animal Breeding and Genomics Centre, Wageningen UR Livestock Research, Wageningen, the Netherlands.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
- Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium.
- Animal Embryo Centre, Maria-Hoop, the Netherlands.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
MeSH Terms
- Animals
- Cryopreservation / veterinary
- Dimethyl Sulfoxide / administration & dosage
- Dose-Response Relationship, Drug
- Embryo Culture Techniques
- Embryo Transfer
- Female
- Glycerol / administration & dosage
- Horses / embryology
- In Vitro Oocyte Maturation Techniques / veterinary
- Oocytes / physiology
- Pregnancy
- Tissue Preservation / methods
- Tissue Preservation / veterinary
- Vitrification
Citations
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