FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Reproduction in domestic animals = Zuchthygiene2021; 56(10); 1358-1362; doi: 10.1111/rda.13990

Evaluating the use of piezo manipulator, laser or their combination for blastocoel cavity puncture to improve cryopreservation outcomes of large equine embryos.

Abstract: The main difficulty of large equine embryo cryopreservation is the replacement of blastocoel fluid with cryoprotectant solution. The objective of this study was to improve the cryopreservation of large equine embryos with PMAP and/or LAP. Embryos were collected via the non-surgical transcervical procedure and divided into three groups based on their size (A ≤ 300 µm, 300 µm<B 300 µm). However, more research is required to find the best method for embryos ≥700 µm.
© 2021 Wiley-VCH GmbH.
Get Full Text
Publication Date: 2021-07-17 PubMed ID: 34254367DOI: 10.1111/rda.13990Google 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.
LinkedInCopy
  • 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.

This study investigates methods to improve the preservation of large horse embryos by using a piezo manipulator, laser, or a combination of both to replace fluid in the egg cell with a cryopreservative solution. Although the techniques provided positive pregnancy results for embryos up to 700µm in size, further research is necessary for embryos larger than this.

Objectives and Methods of the Study

  1. The primary aim of this study was to enhance the cryopreservation process of large horse embryos. The overall process involves the replacement of blastocoel fluid with a concoction known as cryoprotectant.
  2. The researchers collected the embryos through a non-invasive transcervical procedure. These embryos were then categorized into three distinct groups according to their size: Group A, with sizes up to 300µm; Group B, sizes ranging from 300µm to 700µm; and Group C, sizes above 700µm.
  3. The piezo manipulator (a precision instrument used to modify a microscopic specimen), the laser pulse, or a combination were used to puncture six chosen embryos.

Cryopreservation, Cultivation and Transfer

  1. Post-puncture, all embryos underwent cryopreservation on a Cryotop®, a simple and efficient tool used to vitrify biological materials at ultra-rapid cooling rates.
  2. After being frozen and subsequently thawed, the embryos were cultured for three hours before their transfer into the recipient mares.

Pregnancy Diagnosis and Findings

  1. One week after the transfer, pregnancy was confirmed by executing an ultrasound.
  2. Out of the six implanted embryos, two resulted in a pregnancy. For Group A and B embryos, there were positive pregnancy results; however, for those in Group C (≥700µm), the results were negative.
  3. Laser-assisted puncture thus proved beneficial in extracting blastocoel fluid and replacing it with cryoprotectant.

Limitations and Further Research

  1. The first successful pregnancy using laser puncture-assisted frozen-thawed horse embryo larger than 300µm was reported in this study.
  2. Although the study showed promising results, the embryos that were 700µm or larger failed to result in a positive pregnancy, indicating a need for further research to determine the most effective method for cryopreserving larger embryos.

Cite This Article

APA
Rajabi-Toustani R, Watanabe H, Tsogtgerel M, Gao Y, Canbo L, Haneda S, Cheong SH, Nambo Y. (2021). Evaluating the use of piezo manipulator, laser or their combination for blastocoel cavity puncture to improve cryopreservation outcomes of large equine embryos. Reprod Domest Anim, 56(10), 1358-1362. https://doi.org/10.1111/rda.13990

Publication

Reproduction in domestic animals = Zuchthygiene
ISSN: 1439-0531
NlmUniqueID: 9015668
Country: Germany
Language: English
Volume: 56
Issue: 10
Pages: 1358-1362

Researcher Affiliations

Rajabi-Toustani, Reza
  • Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
Watanabe, Hiroyuki
  • Department of Life and Food Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
Tsogtgerel, Munkhtuul
  • Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
  • United Graduate School of Veterinary Sciences, Gifu University, Gifu, Japan.
Gao, Yuanzhi
  • Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
Canbo, Li
  • Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
Haneda, Shingo
  • Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
Cheong, Soon Hon
  • Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
Nambo, Yasuo
  • Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
  • United Graduate School of Veterinary Sciences, Gifu University, Gifu, Japan.

MeSH Terms

  • Animals
  • Blastocyst / physiology
  • Cryopreservation / methods
  • Cryopreservation / veterinary
  • Cryoprotective Agents
  • Embryo Transfer / instrumentation
  • Embryo Transfer / methods
  • Embryo Transfer / veterinary
  • Embryo, Mammalian
  • Female
  • Horses / embryology
  • Lasers
  • Micromanipulation / methods
  • Micromanipulation / veterinary
  • Pregnancy

Grant Funding

  • Livestock Promotional Subsidy from the Japan Racing Association

References

This article includes 13 references
  1. Allen WR, Stewart F. Equine placentation. Reproduction, Fertility and Development 13(7-8), 623-634.
    doi: 10.1071/rd01063google scholar: lookup
  2. Choi YH, Velez IC, Riera FL, Roldán JE, Hartman DL, Bliss SB, Blanchard TL, Hayden SS, Hinrichs K. Successful cryopreservation of expanded equine blastocysts. Theriogenology 76(1), 143-152.
    doi: 10.1016/j.theriogenology.2011.01.028google scholar: lookup
  3. Davidson LM, Liu Y, Griffiths T, Jones C, Coward K. Laser technology in the ART laboratory: A narrative review. Reproductive BioMedicine Online 38, 725-739.
    doi: 10.1016/j.rbmo.2018.12.011google scholar: lookup
  4. Diaz F, Bondiolli K, Paccamonti D, Gentry GT. Cryopreservation of Day 8 equine embryos after blastocyst micromanipulation and vitrification. Theriogenology 85(5), 894-903.
    doi: 10.1016/j.theriogenology.2015.10.039google scholar: lookup
  5. Eldridge-Panuska WD, Caracciolo Di Brienza V, Seidel GE, Squires EL, Carnevale EM. Establishment of pregnancies after serial dilution or direct transfer by vitrified equine embryos. Theriogenology 63(5), 1308-1319.
    doi: 10.1016/j.theriogenology.2004.06.015google scholar: lookup
  6. Hannan MA, Haneda S, Itami Y, Wachi S, Saitoh T, Cheong SH, Nambo Y. Successful embryo transfer from Hokkaido native pony after artificial insemination with frozen semen. Journal of Veterinary Medical Science 81(2), 241-244.
    doi: 10.1292/jvms.18-0414google scholar: lookup
  7. Hannan MA, Haneda S, Murata K, Takeuchi S, Cheong SH, Nambo Y. Birth of first foals through embryo transfer after artificial insemination using frozen semen in Japan. Journal of Reproduction and Development 66(2), 193-197.
    doi: 10.1262/jrd.2019-117google scholar: lookup
  8. Mukaida T, Oka C, Goto T, Takahashi K. Artificial shrinkage of blastocoeles using either a micro-needle or a laser pulse prior to the cooling steps of vitrification improves survival rate and pregnancy outcome of vitrified human blastocysts. Human Reproduction 21(12), 3246-3252.
    doi: 10.1093/humrep/del285google scholar: lookup
  9. Scherzer J, Davis C, Hurley D. Laser-assisted vitrification of large equine embryos. Reproduction in Domestic Animals 46(6), 1104-1106.
    doi: 10.1111/j.1439-0531.2011.01795.xgoogle scholar: lookup
  10. Stout T. Cryopreservation of equine embryos: Current state-of-the-art. Reproduction in Domestic Animals 47(Suppl 3), 84-89.
    doi: 10.1111/j.1439-0531.2012.02030.xgoogle scholar: lookup
  11. Stout TAE, Meadows S, Allen WR. Stage-specific formation of the equine blastocyst capsule is instrumental to hatching and to embryonic survival in vivo. Animal Reproduction Science 87(3-4), 269-281.
    doi: 10.1016/j.anireprosci.2004.11.009google scholar: lookup
  12. Tremoleda JL, Stout TAE, Lagutina I, Lazzari G, Bevers MM, Colenbrander B, Galli C. Effects of in vitro production on horse embryo morphology, cytoskeletal characteristics, and blastocyst capsule formation. Biology of Reproduction 69(6), 1895-1906.
    doi: 10.1095/biolreprod.103.018515google scholar: lookup
  13. Wilsher S, Rigali F, Kovacsy S, Allen W. Puncture of the equine embryonic capsule and its repair in vivo and in vitro. Journal of Equine Veterinary Science 93, 103194.
    doi: 10.1016/j.jevs.2020.103194google scholar: lookup

Citations

This article has been cited 2 times.
  1. Rajabi-Toustani R, Tsogtgerel M, Gao Y, Li C, Sakato M, Haneda S, Cheong SH, Nambo Y. First Kiso pony foal produced via transfer of long-distance shipped fresh embryo to Hokkaido native pony. J Reprod Dev 2023 Apr 3;69(2):125-128.
    doi: 10.1262/jrd.2022-125pubmed: 36724993google scholar: lookup
  2. Hannan MA, Watanabe H, Takeyama A, Yoshida S, Wudamu D, Lkhagvasuren N, Claes A, Stout TAE, Cheong SH, Haneda S, Nambo Y. In vitro embryo production via ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) in pure and crossbred Japanese Hokkaido native ponies. J Reprod Dev 2025 Jun 6;71(3):191-194.
    doi: 10.1262/jrd.2025-011pubmed: 40350303google scholar: lookup
Subscribe to our newsletter
Join 99,780 horse owners like you who receive our email updates on horse health and nutrition.