Omneity® Pellets
All-In-One Vitamin & Mineral Pellet
In vitro embryo production via ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) in pure and crossbred Japanese Hokkaido native ponies.
Abstract: This study evaluated the viability of in vitro embryo production using ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) as breeding techniques for pure and crossbred Hokkaido native ponies (n = 9). Oocytes were collected using transvaginal ultrasound-guided follicle aspiration. ICSI was performed on in vitro matured oocytes using frozen semen. Embryonic cultures were monitored using time-lapse cinematography. Blastocysts were cryopreserved and, after thawing, were transferred non-surgically into recipient mares. Over nine OPU sessions, the mean number of aspirated follicles was 23.9 (range, 13-49). The oocyte recovery and maturation rates were 35.3% (76/215) and 61.5% (40/65), respectively. The cleavage rate was 57.5% (23/40). Of cleaved embryos, 56.5% (13/23) were arrested at the 4-cell to 8-cell stage, and five developed into early-blastocyst. Three embryos were transferred, resulting in a successful pregnancy. In conclusion, OPU-ICSI is a viable assisted reproductive technology for enhancing the population of Japanese native horses.
Publication Date: 2025-05-10 PubMed ID: 40350303PubMed Central: PMC12151631DOI: 10.1262/jrd.2025-011Google 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.
- 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 aims to assess the effectiveness of in vitro embryo production using ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) techniques in breeding Japanese Hokkaido native ponies.
Methodology
- The study involved 9 pure and crossbred Hokkaido native ponies.
- Oocytes, or female egg cells, were collected via the transvaginal ultrasound-guided follicle aspiration technique, also known as ovum pick-up (OPU).
- These collected oocytes were then matured in vitro, that is, outside the living organism in a controlled lab setting.
- Intracytoplasmic sperm injection (ICSI) was then performed on these matured oocytes. In ICSI, a single sperm cell is injected directly into the cytoplasm of the egg using a microneedle. The sperm used for the study was frozen or cryopreserved.
Embryo Culture and Transfer
- The resulting embryo cultures were monitored using time-lapse cinematography, which provides detailed and continuous visual tracking of embryo development.
- The developed blastocysts, an early stage embryo, were then cryopreserved for future use. Cryopreservation is the process of preserving cells or whole tissues by cooling them to sub-zero temperatures.
- Post-thawing, these blastocysts were non-surgically transferred into recipient mares.
Results
- The research showed that on average, 23.9 follicles could be aspirated in each of the nine OPU sessions, with the number ranging from 13 to 49.
- The oocyte recovery rate was 35.3% (76 out of 215), indicating that about a third of the oocytes could be successfully retrieved from the aspirated follicles.
- The oocyte maturation rate was 61.5% (40 out of 65), showing that about two-thirds of the retrieved oocytes could be matured successfully in vitro.
- The cleavage rate, or the rate at which the fertilized oocytes started dividing into multiple cells, was 57.5% (23 out of 40).
- Of these cleaved embryos, about 56.5% (13 out of 23) were arrested at the 4-cell to 8-cell stage, that is, their development ceased at this stage.
- However, five embryos still managed to develop into early blastocysts.
- Three of these resulting embryos were successfully transferred leading to a successful pregnancy.
Conclusion
The study concluded that the use of OPU-ICSI is a viable assisted reproductive technology for boosting the population of Japanese native horses. The method demonstrated a fair success rate in terms of oocyte recovery, in vitro maturation, and resulting pregnancies.
Cite This Article
APA
Hannan MA, Watanabe H, Takeyama A, Yoshida S, Wudamu D, Lkhagvasuren N, Claes A, Stout TAE, Cheong SH, Haneda S, Nambo Y.
(2025).
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, 71(3), 191-194.
https://doi.org/10.1262/jrd.2025-011 Publication
Researcher Affiliations
- Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan.
- Department of Physiology, Faculty of Veterinary, Animal and Biomedical Sciences, Khulna Agricultural University, 9202, Khulna, Bangladesh.
- Department of Life and Food Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, Japan.
- Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan.
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584CM Utrecht, the Netherlands.
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584CM Utrecht, the Netherlands.
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, NY 14853, USA.
- Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Sciences, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan.
MeSH Terms
- Animals
- Sperm Injections, Intracytoplasmic / veterinary
- Sperm Injections, Intracytoplasmic / methods
- Horses / embryology
- Female
- Pregnancy
- Male
- Embryo Culture Techniques / veterinary
- Embryo Transfer / veterinary
- Fertilization in Vitro / veterinary
- Fertilization in Vitro / methods
- Oocyte Retrieval / veterinary
- Cryopreservation / veterinary
- Embryonic Development
- Blastocyst / cytology
- Japan
- Oocytes / cytology
- Breeding / methods
- In Vitro Oocyte Maturation Techniques / veterinary
Conflict of Interest Statement
None of the authors have any financial or personal relationships that could influence or bias the content of this paper.
References
This article includes 19 references
- Dell’Aquila ME, De Felici M, Massari S, Maritato F, Minoia P. Effects of fetuin on zona pellucida hardening and fertilizability of equine oocytes matured in vitro.. Biol Reprod 1999; 61: 533–540.
- Alm H, Torner H, Blottner S, Nürnberg G, Kanitz W. Effect of sperm cryopreservation and treatment with calcium ionophore or heparin on in vitro fertilization of horse oocytes.. Theriogenology 2001; 56: 817–829.
- Galli C, Colleoni S, Duchi R, Lagutina I, Lazzari G. Developmental competence of equine oocytes and embryos obtained by in vitro procedures ranging from in vitro maturation and ICSI to embryo culture, cryopreservation and somatic cell nuclear transfer.. Anim Reprod Sci 2007; 98: 39–55.
- Tremoleda JL, Stout TA, 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.. Biol Reprod 2003; 69: 1895–1906.
- Squires E, Wilson J, Kato H, Blaszczyk A. A pregnancy after intracytoplasmic sperm injection into equine oocytes matured in vitro.. Theriogenology 1996; 45: 306.
- Galli C, Crotti G, Turini P, Duchi R, Mari G, Zavaglia G, Duchamp G, Daels P, Lazzari G. Frozen- thawed embryos produced by Ovum Pick up of immature oocytes and ICSI are capable to establish pregnancies in the horse.. Theriogenology 2002; 58: 705–708.
- Galli C, Colleoni S, Duchi R, Lazzari G. Male factors affecting the success of equine in vitro embryo production by ovum pickup-intracytoplasmic sperm injection in a clinical setting.. J Equine Vet Sci 2016; 43: S6–S10.
- Galli C, Duchi R, Colleoni S, Lagutina I, Lazzari G. Ovum pick up, intracytoplasmic sperm injection and somatic cell nuclear transfer in cattle, buffalo and horses: from the research laboratory to clinical practice.. Theriogenology 2014; 81: 138–151.
- Galli C, Colleoni S, Claes A, Beitsma M, Deelen C, Necchi D, Duchi R, Lazzari G, Stout T. Overnight shipping of equine oocytes from remote locations to an ART laboratory enables access to the flexibility of ovum pick up-ICSI and embryo cryopreservation technologies.. J Equine Vet Sci 2016; 41: 82.
- Claes A, Cuervo-Arango J, van den Broek J, Galli C, Colleoni S, Lazzari G, Deelen C, Beitsma M, Stout TA. Factors affecting the likelihood of pregnancy and embryonic loss after transfer of cryopreserved in vitro produced equine embryos.. Equine Vet J 2019; 51: 446–450.
- Brooks KE, Daughtry BL, Metcalf E, Masterson K, Battaglia D, Gao L, Park B, Chavez SL. Assessing equine embryo developmental competency by time-lapse image analysis.. Reprod Fertil Dev 2019; 31: 1840–1850.
- Meyers S, Burruel V, Kato M, de la Fuente A, Orellana D, Renaudin C, Dujovne G. Equine non-invasive time-lapse imaging and blastocyst development.. Reprod Fertil Dev 2019; 31: 1874–1884.
- Martino NA, Marzano G, Mastrorocco A, Lacalandra GM, Vincenti L, Hinrichs K, Dell Aquila ME. Use of time-lapse imaging to evaluate morphokinetics of in vitro equine blastocyst development after oocyte holding for two days at 15°C versus room temperature before intracytoplasmic sperm injection.. Reprod Fertil Dev 2019; 31: 1862–1873.
- Lewis N, Schnauffer K, Hinrichs K, Morganti M, Troup S, Argo C. Morphokinetics of early equine embryo development in vitro using time-lapse imaging, and use in selecting blastocysts for transfer.. Reprod Fertil Dev 2019; 31: 1851–1861.
- Goszczynski DE, Tinetti PS, Choi YH, Hinrichs K, Ross PJ. Genome activation in equine in vitro-produced embryos.. Biol Reprod 2022; 106: 66–82.
- Lazzari G, Colleoni S, Crotti G, Turini P, Fiorini G, Barandalla M, Landriscina L, Dolci G, Benedetti M, Duchi R, Galli C. Laboratory production of equine embryos.. J Equine Vet Sci 2020; 89: 103097.
- Toyoda Y, Yokoyama M, Hosi T. Studies on the fertilization of mouse eggs in vitro: I. In vitro fertilization of eggs by fresh epididymal sperm (in Japanese).. Jpn J Anim Reprod 1971; 16: 147–151.
- Rajabi-Toustani R, Watanabe H, Tsogtgerel M, Gao Y, Canbo L, Haneda S, Cheong SH, Nambo Y. 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 2021; 56: 1358–1362.
- 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.. J Vet Med Sci 2019; 81: 241–244.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
