J Vet Sci.  2015 Dec;16(4):509-516. 10.4142/jvs.2015.16.4.509.

Cloned foal derived from in vivo matured horse oocytes aspirated by the short disposable needle system

Affiliations
  • 1Department of Theriogenology, College of Veterinary Medicine and the Research Institute for Veterinary Science, Seoul National University, Seoul 08826, Korea. snujang@snu.ac.kr
  • 2Department of Surgery, College of Veterinary Medicine and the Research Institute for Veterinary Science, Seoul National University, Seoul 08826, Korea.
  • 3Department of Life Science, Sogang University, Seoul 04107, Korea.
  • 4College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea. scyeon@gnu.ac.kr
  • 5Emergence Center for Food-Medicine Personalized Therapy System, Advanced Institutes of Convergence Technology, Seoul National University, Suwon 16229, Korea.
  • 6Farm Animal Clinical Training and Research Center, Institutes of GreenBio Science Technology, Seoul National University, Pyeongchang 25354, Korea.

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

Keyword

cloned horse; embryo transfer; follicle aspiration; short needle; somatic cell nuclear transfer

MeSH Terms

Clone Cells*
DNA
DNA, Mitochondrial
Embryo Transfer
Embryology
Embryonic Structures
Horses*
Humans
Mental Competency
Microsatellite Repeats
Needles*
Oocyte Retrieval
Oocytes*
Tissue Donors
DNA
DNA, Mitochondrial

Figure

  • Fig. 1 Representative pictures of various equine oocytes. (A) Ex-cumulus oocyte complexes (COCs) with well-expanded cumulus or mucous. (B) COCs with cumulus or corona cells tightly surrounding oocytes. (C) COCs with only corona radiata or partial layer of cumulus. (D) COCs with ruptured cytoplasm. (E) Oocytes without the first polar body. (F) Oocytes with degenerated cytoplasm.

  • Fig. 2 Cloned foal (left) and surrogate mother (right).


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