Lee Wonyou, Song Kilyoung, Lee Inhyung, Shin Hyungdo, Lee Byeong Chun, Yeon Seongchan, Jang Goo
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.
J Vet Sci. 2015;16(4):509-16. doi: 10.4142/jvs.2015.16.4.509.
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.
经阴道超声引导下卵泡抽吸术是获取受体卵母细胞用于马体细胞克隆(SCNT)的一种方法。本研究旨在:(1)通过比较短的一次性针头系统(14-G)与长的双腔针头(12-G)的卵母细胞回收率,评估从排卵前卵泡中抽吸卵母细胞的可能性;(2)研究体细胞克隆和胚胎移植后回收的卵母细胞的发育能力。短的一次性针头与长针头的回收率无显著差异(分别为47.5%和35.0%)。26个体细胞克隆胚胎被移植到13匹母马体内,其中一匹母马在第342天分娩出一头活仔。对微卫星DNA进行分析后发现,克隆马驹与细胞供体完全匹配,克隆马驹的线粒体DNA与卵母细胞供体的线粒体DNA相同。这些结果表明,短的一次性针头系统可用于回收卵母细胞,作为体细胞克隆的细胞质供体,用于克隆马驹的生产以及马胚胎学的其他应用。