Cloned foal derived from in vivo matured horse oocytes aspirated by the short disposable needle system.
Abstract: Transvaginal ultrasound-guided follicle aspiration is one method of obtaining recipient oocytes for equine somatic cell nuclear transfer (SCNT). This study was conducted: (1) to evaluate the possibility of oocyte aspiration from pre-ovulatory follicles using a short disposable needle system (14-G) by comparing the oocyte recovery rate with that of a long double lumen needle (12-G); (2) to investigate the developmental competence of recovered oocytes after SCNT and embryo transfer. The recovery rates with the short disposable needle vs. the long needle were not significantly different (47.5% and 35.0%, respectively). Twenty-six SCNT embryos were transferred to 13 mares, and one mare delivered a live offspring at Day 342. There was a perfect identity match between the cloned foal and the cell donor after analysis of microsatellite DNA, and the mitochondrial DNA of the cloned foal was identical with that of the oocyte donor. These results demonstrated that the short disposable needle system can be used to recover oocytes to use as cytoplasts for SCNT, in the production of cloned foals and for other applications in equine embryology.
Publication Date: 2015-06-30 PubMed ID: 26119166PubMed Central: PMC4701744DOI: 10.4142/jvs.2015.16.4.509Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research study conducted experiments to examine the feasibility of using a short disposable needle system for obtaining horse oocytes, or female reproductive cells, for replication through somatic cell nuclear transfer (SCNT). The authors found this process to be just as effective as using a long needle, and managed to produce a cloned foal using the harvested oocytes.
Techniques and Methodology
- The authors used transvaginal, ultrasound-assisted follicle aspiration to obtain recipient oocytes for cloning through SCNT. This technique allows for the collection of mature oocytes from the ovaries of female horses (mares).
- The researchers compared the oocyte recovery rate of a short disposable needle system with that of a traditional long needle. They found that the rates were not significantly different, with the short disposable needle retrieving 47.5% of oocytes and the long needle retrieving 35.0%.
Somatic Cell Nuclear Transfer (SCNT) and Embryo Transfer
- The harvested oocytes were then used in SCNT to replicate genetic material.
- Twenty-six of the embryos created through SCNT were transferred to 13 different mares for gestation. One of these mares successfully gave birth to a cloned foal at 342 days, demonstrating the viability of the embryos produced through this technique.
DNA Analysis and Results
- The researchers used microsatellite DNA analysis to compare the genetic information of the cloned foal with that of its cell donor. They found an exact genetic match between the foal and the donor, confirming the success of the cloning process.
- The researchers also compared the mitochondrial DNA of the foal with that of the oocyte donor. Again, the genetic material was identical, showing that the short needle aspiration method effectively collected oocytes suitable for SCNT.
Conclusion
- Based on these results, the authors concluded that the short disposable needle system can be effectively used to collect oocytes for cloning purposes in horses.
- The research further establishes the potential of SCNT as a viable technique for cloning horses and possibly other large, domestic animals.
- Finally, this system might have important implications for producing cloned animals in veterinary medicine and for preserving endangered equine species.
Cite This Article
APA
Lee W, Song K, Lee I, Shin H, Lee BC, Yeon S, Jang G.
(2015).
Cloned foal derived from in vivo matured horse oocytes aspirated by the short disposable needle system.
J Vet Sci, 16(4), 509-516.
https://doi.org/10.4142/jvs.2015.16.4.509 Publication
Researcher Affiliations
- Department of Theriogenology, College of Veterinary Medicine and the Research Institute for Veterinary Science, Seoul National University, Seoul 08826, Korea.
- Department of Theriogenology, College of Veterinary Medicine and the Research Institute for Veterinary Science, Seoul National University, Seoul 08826, Korea.
- Department of Surgery, College of Veterinary Medicine and the Research Institute for Veterinary Science, Seoul National University, Seoul 08826, Korea.
- Department of Life Science, Sogang University, Seoul 04107, Korea.
- Department of Theriogenology, College of Veterinary Medicine and the Research Institute for Veterinary Science, Seoul National University, Seoul 08826, Korea.
- College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea.
- Department of Theriogenology, College of Veterinary Medicine and the Research Institute for Veterinary Science, Seoul National University, Seoul 08826, Korea.
- Emergence Center for Food-Medicine Personalized Therapy System, Advanced Institutes of Convergence Technology, Seoul National University, Suwon 16229, Korea.
- Farm Animal Clinical Training and Research Center, Institutes of GreenBio Science Technology, Seoul National University, Pyeongchang 25354, Korea.
MeSH Terms
- Animals
- Cloning, Organism / veterinary
- DNA, Mitochondrial / genetics
- Embryo Transfer / veterinary
- Embryo, Mammalian
- Female
- Horses
- Microsatellite Repeats
- Needles / veterinary
- Nuclear Transfer Techniques / veterinary
- Oocyte Retrieval / veterinary
- Ovarian Follicle / cytology
- Pregnancy
Conflict of Interest Statement
There is no conflict of interest.
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Citations
This article has been cited 2 times.- Olivera R, Moro LN, Jordan R, Pallarols N, Guglielminetti A, Luzzani C, Miriuka SG, Vichera G. Bone marrow mesenchymal stem cells as nuclear donors improve viability and health of cloned horses. Stem Cells Cloning 2018;11:13-22.
- Olivera R, Moro LN, Jordan R, Luzzani C, Miriuka S, Radrizzani M, Donadeu FX, Vichera G. In Vitro and In Vivo Development of Horse Cloned Embryos Generated with iPSCs, Mesenchymal Stromal Cells and Fetal or Adult Fibroblasts as Nuclear Donors. PLoS One 2016;11(10):e0164049.
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