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Cryopreservation of immature equine oocytes, comparing a solid surface vitrification process with open pulled straws and the use of a synthetic ice blocker.
Abstract: The objective was to evaluate the effect of three cryopreservation methods on the in vitro maturation (IVM) and membrane integrity (MIn) of immature equine oocytes. An open pulled straw (OPS) method, a novel solid surface vitrification (SSV) process, and the addition of a synthetic ice blocker were evaluated. Compared with the control group (N=269), the OPS (N=159) and the SSV (N=202) cryopreservation methods decreased both IVM (50.9 vs. 13.3 and 9.4%, respectively; P<0.001) and MIn (76.6 vs. 31.1 and 33.7%; P<0.001) of immature equine oocytes. However, inclusion of 0.1% ice blocker in the OPS vitrification process increased the rates of both IVM (30.5%; P<0.01) and MIn (45.8%; P<0.05) of the oocytes (N=59). Including 0.1% ice blocker in the SSV process improved the IVM rate (20.9%; P<0.05), whereas MIn remained compromised in this group (N=67). However, increasing the concentration of the ice blocker (to 1.0%) in the cryopreservation methods did not significantly improve rates of IVM. In conclusion, the addition of a synthetic ice blocker (0.1%) to both cryopreservation processes significantly increased rates of both IVM and MIn of immature equine oocytes cryopreserved by OPS.
Copyright © 2012 Elsevier Inc. All rights reserved.
Publication Date: 2011-08-10 PubMed ID: 21835449DOI: 10.1016/j.theriogenology.2011.07.008Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Comparative Study
- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
- Animal Science
- Assisted Reproductive Techniques
- Biotechnology
- Cell Viability
- Cells
- Cellular Membranes
- Comparative Study
- Cryopreservation
- Equine Diseases
- Equine Health
- Equine Science
- Experimental Methods
- Freezing Technique
- In Vitro Research
- Oocyte
- Physiology
- Reproductive Technology
- Veterinary Medicine
- Veterinary Research
Summary
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This research compares different methods of preserving immature equine oocytes or horse eggs. The study primarily focuses on the impact of the Open Pulled Straw method, Solid Surface Vitrification, and the use of a synthetic ice blocker. Results showed that a synthetic ice blocker increased the rates of in vitro maturation and membrane integrity in the eggs preserved.
Comparing Cryopreservation Methods
- The goal of the experiment was to compare the effects of three different cryopreservation methods on immature equine oocytes (horse eggs). The main points of comparison were the rates of in vitro maturation (IVM) and membrane integrity (MIn) of the oocytes following cryopreservation.
- Open Pulled Straw (OPS), Solid Surface Vitrification (SSV), and the addition of a synthetic ice blocker were the different methods used for cryopreservation in this study.
- The control group consisted of non-cryopreserved eggs (N=269). For the experimental groups, OPS was used on 159 cells and SSV on 202 cells.
- Compared to the control group, both OPS and SSV methods resulted in significantly lower rates of IVM and MIn in the cryopreserved cells. These decreases are statistically significant as indicated by the P values of less than 0.001.
Use of Synthetic Ice Blocker
- The use of a synthetic ice blocker in the cryopreservation process significantly increased the success of both IVM and MIn in the OPS method, as demonstrated by P values less than 0.01 and 0.05 respectively.
- Adding a synthetic ice blocker to the SSV process increased the IVM rates but did not significantly improve the MIn. This indicates that while the eggs matured, their membrane integrity was not preserved as well.
- The effect of the ice blocker was not improved by increasing its concentration beyond 0.1%. This suggests an optimal ice blocker concentration for enhancing both IVM and MIn.
Conclusions
- To conclude, the addition of a synthetic ice blocker (0.1%) significantly improved the outcomes of both IVM and MIn for immature equine oocytes cryopreserved with the OPS method. This could be an essential step for optimizing cryopreservation methods in the future for this species.
Cite This Article
APA
de Leon PM, Campos VF, Corcini CD, Santos EC, Rambo G, Lucia T, Deschamps JC, Collares T.
(2011).
Cryopreservation of immature equine oocytes, comparing a solid surface vitrification process with open pulled straws and the use of a synthetic ice blocker.
Theriogenology, 77(1), 21-27.
https://doi.org/10.1016/j.theriogenology.2011.07.008 Publication
Researcher Affiliations
- Laboratório de Embriologia Molecular e Transgênese, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
MeSH Terms
- Animals
- Cell Survival
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents / pharmacology
- Fertilization in Vitro / veterinary
- Horses
- Oocytes / cytology
- Oocytes / drug effects
- Oocytes / ultrastructure
Citations
This article has been cited 8 times.- 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).
- Choi HW, Jang H. Application of Nanoparticles and Melatonin for Cryopreservation of Gametes and Embryos. Curr Issues Mol Biol 2022 Sep 5;44(9):4028-4044.
- Guo Z, Chen W, Lv L, Liu D. Meta-analysis of melatonin treatment and porcine somatic cell nuclear transfer embryo development. Anim Reprod 2021;18(3):e20210031.
- Angel-Velez D, De Coster T, Azari-Dolatabad N, Fernandez-Montoro A, Benedetti C, Bogado Pascottini O, Woelders H, Van Soom A, Smits K. New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification. Animals (Basel) 2021 Oct 28;11(11).
- Zhao X, Wang D, Wu Z, Pan B, Yang H, Zeng C, Zhang M, Liu G, Han H, Zhou G. Female Reproductive Performance in the Mouse: Effect of Oral Melatonin. Molecules 2018 Jul 25;23(8).
- Li W, Cheng K, Zhang Y, Meng Q, Zhu S, Zhou G. No effect of exogenous melatonin on development of cryopreserved metaphase II oocytes in mouse. J Anim Sci Biotechnol 2015;6(1):42.
- Rimayanti R, Khairullah AR, Madyawati SP, Rantam FA, Mustofa I, Widjiati W, Srianto P, Akintunde AO, Pratama BP, Ahmad RZ, Wardhani BWK, Khalisa AT, Wibowo S. The role of heat shock protein 70 on oocyte apoptosis during vitrification. Open Vet J 2025;15(10):4847-4864.
- Maclellan LJ, Albertini DF, Stokes JE, Carnevale EM. Use of confocal microscopy and intracytoplasmic sperm injection (ICSI) to assess viability of equine oocytes from young and old mares after vitrification. J Assist Reprod Genet 2023 Nov;40(11):2565-2576.
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