Wolf Don P
Division of Reproductive Sciences, Oregon National Primate Research Center Beaverton, Oregon, USA.
Reprod Biol Endocrinol. 2004 Jun 16;2:37. doi: 10.1186/1477-7827-2-37.
The assisted reproductive technologies (ARTs) have been used in the production of rhesus monkey offspring at the Oregon National Primate Research Center (ONPRC) and that experience is summarized here. Additionally these technologies serve as a source of oocytes/embryos for monozygotic twinning, embryonic stem (ES) cell derivation and cloning. High fertilization efficiencies were realized with conventional insemination or following the use of intracytoplasmic sperm injection (ICSI) and approximately 50% of the resulting embryos grew in vitro to blastocysts. Both fresh and frozen sperm were employed in fertilization by ICSI and the resulting embryos could be low temperature stored for subsequent thawing and transfer when a synchronized recipient female was available or after shipment to another facility. Following the transfer of up to 3 embryos, an overall pregnancy rate of 30% was achieved with increasing rates dependent upon the number of embryos transferred. Singleton pregnancy outcomes following the transfer of ART produced embryos were similar to those observed in a control group of animals in the timed mated breeding colony at ONPRC. ICSI produced embryos were used in efforts to create monozygotic twins by blastomere separation or blastocyst splitting. While pregnancies were achieved following the transfer of demi-embryos, only one was a twin and it was lost to spontaneous abortion. ICSI produced embryos have also served as the source of blastocysts for the derivation of embryonic stem cells. These pluripotent cells hold potential for cell based therapies and we consider the monkey an important translational model in which to evaluate safety, efficacy and feasibility of regenerative medicine approaches based on the transplantation of stem cell-derived progeny. Finally, efforts to produce genetically-identical monkeys by nuclear transfer have been briefly summarized.
俄勒冈国家灵长类动物研究中心(ONPRC)已将辅助生殖技术(ARTs)用于恒河猴后代的生产,本文总结了这一经验。此外,这些技术还为单卵孪生、胚胎干细胞(ES)衍生和克隆提供卵母细胞/胚胎来源。常规授精或使用胞浆内单精子注射(ICSI)后实现了高受精效率,约50%的所得胚胎在体外发育至囊胚。ICSI受精采用了新鲜和冷冻精子,所得胚胎可低温保存,以便在有同步受体雌性时随后解冻和移植,或在运往另一机构后进行解冻和移植。移植多达3个胚胎后,总体妊娠率达到30%,妊娠率随移植胚胎数量的增加而提高。ART生产的胚胎移植后的单胎妊娠结局与ONPRC定时交配繁殖群体中对照组动物观察到的结局相似。ICSI生产的胚胎被用于通过卵裂球分离或囊胚分割来创造单卵双胞胎。虽然半胚胎移植后实现了妊娠,但只有一例是双胞胎,且因自然流产而丢失。ICSI生产的胚胎也作为囊胚来源用于胚胎干细胞的衍生。这些多能细胞具有细胞治疗的潜力,我们认为猴子是评估基于干细胞衍生后代移植的再生医学方法的安全性、有效性和可行性的重要转化模型。最后,简要总结了通过核移植生产基因相同猴子的努力。