Wolfgang M J, Eisele S G, Knowles L, Browne M A, Schotzko M L, Golos T G
Wisconsin Regional Primate Research Center, University of Wisconsin Medical School, Madison 53715-1299, USA.
J Med Primatol. 2001 Jun;30(3):148-55. doi: 10.1111/j.1600-0684.2001.tb00003.x.
Embryo transfer in the rhesus monkey has been historically limited to transfer of cleavage stage embryos. In order to allow genetic manipulation of rhesus embryos in vitro, without using invasive surgical techniques, it is important to explore the transfer of morula and blastocyst stage embryos. Embryos were produced by in vitro fertilization from gonadotropin-stimulated monkeys, or were obtained by nonsurgical uterine flushing of naturally mated or artificially inseminated females. Nonsurgical transfer was accomplished by inserting a metal guide through the cervix into the uterus, after which a hollow cell sampler was inserted over the guide. The guide was removed and a catheter was inserted containing one to five embryos. Several pregnancies resulted from in vitro- and in vivo-derived blastocysts, and two pregnancies were carried to term resulting in one live birth. Blood samples were collected regularly to monitor plasma levels of chorionic gonadotropin, luteinizing hormone, and progesterone. The recipients received progesterone as a subcutaneous implant or daily injections from the day of transfer. The approach described in this study provides the opportunity to explore transgenic and chimeric models in the monkey by the development of noninvasive methods to transfer late-stage embryos that have been manipulated in vitro.
恒河猴的胚胎移植在历史上一直局限于卵裂期胚胎的移植。为了在不使用侵入性手术技术的情况下对恒河猴胚胎进行体外基因操作，探索桑椹胚和囊胚期胚胎的移植很重要。胚胎通过对促性腺激素刺激的猴子进行体外受精产生，或者通过对自然交配或人工授精的雌性进行非手术子宫冲洗获得。非手术移植是通过将金属导管经宫颈插入子宫，然后将空心细胞采样器套在导管上完成的。移除导管后，插入含有一至五个胚胎的导管。体外和体内来源的囊胚导致了几次妊娠，两次妊娠足月产，产下一个活婴。定期采集血样以监测血浆中绒毛膜促性腺激素、促黄体生成素和孕酮的水平。从移植当天起，受体接受皮下植入或每日注射孕酮。本研究中描述的方法通过开发非侵入性方法来移植体外操作的晚期胚胎，为在猴子中探索转基因和嵌合模型提供了机会。
