Wolf D P, Thormahlen S, Ramsey C, Yeoman R R, Fanton J, Mitalipov S
Division of Reproductive Sciences, Oregon National Primate Research Center, 505 NW 185th Avenue, Beaverton, OR 97006, USA.
Biol Reprod. 2004 Aug;71(2):486-93. doi: 10.1095/biolreprod.103.025932. Epub 2004 Mar 24.
The assisted reproductive technologies (ARTs) as tailored to the production of rhesus monkeys at the Oregon National Primate Research Center (ONPRC) are described. Efficient fertilization of mature oocytes recovered by aspiration from females subjected to follicular stimulation was achieved with fresh or frozen sperm by intracytoplasmic sperm injection (ICSI). Embryo development to the early cleavage stage occurred at high frequency. Cryopreserved embryos showed high postthaw survival and were also transferred in efforts to establish pregnancies. Three methods of transfer were evaluated, two involving embryo placement into the oviduct, laparoscopy and minilaparotomy, and a nonsurgical, transcervical approach that resulted in uterine deposition. Early cleaving embryos (Days 1-4) were transferred into the oviducts of synchronized recipients with optimal results and pregnancy rates of up to 36%. Pregnancy rates were similar when two fresh or frozen embryos were transferred (28- 30%), although more than two embryos had to be thawed to compensate for embryo loss during freeze-thawing. Normal gestational lengths, birth weights, and growth curves were seen with ART-produced infants compared with infants produced by natural mating in the timed mated breeding (TMB) colony at the ONPRC. In 72 singleton pregnancies established following the transfer of ART-produced embryos, the live-birth rate, at 87.5%, was statistically identical to that for the TMB colony. Further development of the ARTs should result in increasing use of these techniques to augment conventional approaches to propagating monkeys, especially those of defined genotypes.
本文描述了俄勒冈国家灵长类动物研究中心(ONPRC)为恒河猴繁殖量身定制的辅助生殖技术(ARTs)。通过卵泡刺激从雌性体内吸出的成熟卵母细胞,利用新鲜或冷冻精子经胞浆内精子注射(ICSI)实现了高效受精。胚胎发育到早期分裂阶段的频率很高。冷冻保存的胚胎解冻后存活率高,并且也进行了移植以尝试建立妊娠。评估了三种移植方法,两种涉及通过腹腔镜和小型剖腹术将胚胎放置到输卵管中,以及一种非手术的经宫颈方法,该方法可使胚胎沉积在子宫内。将早期分裂的胚胎(第1 - 4天)移植到同步受体的输卵管中,效果最佳,妊娠率高达36%。当移植两个新鲜或冷冻胚胎时,妊娠率相似(28 - 30%),尽管为了弥补冻融过程中的胚胎损失,不得不解冻两个以上的胚胎。与ONPRC定时交配繁殖(TMB)群体中自然交配产生的婴儿相比,ART产生的婴儿具有正常的妊娠期长度、出生体重和生长曲线。在移植ART产生的胚胎后建立的72例单胎妊娠中,活产率为87.5%,在统计学上与TMB群体相同。ARTs的进一步发展应会导致越来越多地使用这些技术来补充传统的猴子繁殖方法,特别是对于具有特定基因型的猴子。