Live foals produced from sperm-injected oocytes derived from pregnant mares
Live foals produced from sperm-injected oocytes derived from pregnant mares
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DOI：
10.1016/s0737-0806(98)80504-2
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发表时间：
1998-11-01
影响因子：
1.3
通讯作者：
Godke, RA
中科院分区：
文献类型：
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作者：
Cochran, R;Meintjes, M;Godke, RA
最近，在马身上进行体外受精(IVF)的结果不如预期。与马体外受精相关的各种问题包括：(1)无法收集大量高质量的卵母细胞；(2)与马卵母细胞体外成熟相关的透明带改变；(3)马精子细胞为这些卵母细胞的体外受精准备不当。因此，本研究旨在通过马卵母细胞的精子注射实现受精，并通过这种体外受精程序产生活的后代。采用经阴道超声引导下取母马早期妊娠卵母细胞(孕14~70d)，体外成熟后进行卵胞浆内单精子注射(ICSI)。然后将注射的卵母细胞在加有15%胎牛血清的Tcm-199或P-1(葡萄糖和无磷)培养液中培养48小时。胚胎在两种培养液中的卵裂率不同(Tcm-199组为47%，P-L组为63%)。此外，在ICSI后48小时，四个I级胚胎被手术移植到四匹受体母马(一枚胚胎/母马)的输卵管中，其中三匹妊娠(75%)在ICSI后第16天超声检查显示子宫体中存在胚胎囊泡。6月23日，一只怀孕328天的活母马出生，随后，第二只健康的母马在怀孕319天后出生。据我们所知，这是第一个使用高效、可重复的经阴道超声引导程序对怀孕母马的体外成熟卵母细胞进行体外受精(通过ICSI)而产生活马驹的报告。
Recently, in vitro fertilization (IVF) in the horse has met with less than anticipated results. Various problems associated with equine IVF include: (1) the inability to collect large numbers of good quality oocytes, (2) the alteration of the zona pellucida associated with in vitro maturation of equine oocytes, and (3) the improper preparation of equine sperm cells for IVF of these oocytes. Therefore, this study was conducted to achieve fertilization via sperm injection of equine oocytes and to produce live offspring from this IVF procedure. Oocytes were collected by transvaginal ultrasound-guided oocyte retrieval procedures from early pregnant mares of mixed breeds (day 14 to day 70 of pregnancy) and were matured in vitro and subjected to intracytoplasmic sperm injection (ICSI). Injected oocytes were then cultured for 48 hours in either TCM-199 or P-1 medium(glucose and phosphate-free medium) supplemented with 15% fetal bovine serum. Cleavage rates for embryos cultured in the two culture media were different (47% vs. 63% in TCM-199 and P-l, respectively). Also, four Grade I embryos were surgically transferred into the oviducts of four recipient mares (one embryo/mare) at 48 hours post-ICSI, with three pregnancies (75%) developing as ultrasonically demonstrated by the presence of an embryonic vesicle in the uterine body by day 16 post-ICSI. On June 23rd one live filly was born after 328 days of gestation and subsequently, a second healthy filly was born after 319 days of gestation. To our knowledge, this is the first report of live foals resulting from in vitro fertilization (via ICSI) of in vitro matured oocytes recovered from pregnant mares using an efficient, repeatable transvaginal ultrasound-guided procedure.