Birth of rhesus monkey infant after in vitro fertilization and nonsurgical embryo transfer.

Abstract

The birth of a rhesus monkey resulting from in vitro fertilization is reported. Oocytes recovered at laparoscopy from five gonadotropin-stimulated donors were inseminated in vitro with sperm preincubated with caffeine and dibutyryl cyclic AMP. After insemination, oocytes were cultured for 33-46 hr. Twenty-two embryos were transferred nonsurgically into 11 recipient females. One recipient showed signs of implantation but did not carry to term. A second female became pregnant after receiving one 4-cell and one 6-cell embryo fertilized in vitro. The subsequent course of early pregnancy and embryonic and fetal development were characteristic of normal singleton pregnancies. A healthy term male infant was delivered by Cesarean section 176 days after fertilization. This birth has validated our procedures for in vitro fertilization of rhesus monkey gametes and provides an experimental model for studies of early embryonic development in primates.There is a long-standing need for models using nonhuman primates to supply basic information on fertilization and early embryogenesis. This need has been increased by the clinical applications of in vitro fertilization (IVF) We have devised procedures for IVF and embryo culture using the rhesus monkey (6). In this species, the average duration of the menstrual cycle (28 days), the regulation of folliculogenesis, and the control of ovulation are all very similar to these processes in women (7). Because the rhesus has served as a model for early human development, extensive data on postnatal behavior and development are available (8). Thus, even subtle developmental anomalies could be readily detected if they were to occur in offspring resulting from IVF and ET. We have previously reported normal cleavage development of IVF rhesus oocytes up to the morula stage (6). However, conclusive evidence for normal fertilization and embryogenesis can be obtained only from birth of normal offspring after ET. We now report the birth of a healthy rhesus male infant after nonsurgical (intrauterine) transfer of two IVF embryos to a recipient female. This birth provides validation of the IVF procedures used in our laboratory and offers an experimental model for studies on fertilization and embryonic development in primates. MATERIALS AND METHODSAll female monkeys used in this study were housed separately from males. Procedures for recovery of oocytes from gonadotropin-stimulated rhesus monkeys, preparation of spermatozoa, IVF, and embryo culture were all as previously described (6). Five female rhesus monkeys were each given a total of 2150 international units of pregnant mare serum gonadotropin (PMSG) from day 4 of the menstrual cycle (day 1 = first day of menses) through day 15. On day 16, 4000 international units of human chorionic gonadotropin was given, and aspiration of follicles during laparoscopy was performed 30 hr later. Oocytes were incubated individually in drops of culture medium TALP (9). After 5-7 hr, oocyte drops (90 Al) were inseminated with 10 1.l of washed ejaculated ...

Transgenic rhesus monkeys produced by gene transfer into early-cleavage–stage embryos using a simian immunodeficiency virus-based vector

“…Fifteen embryos were transferred into five pregnant monkeys, who gave birth to four healthy offspring. These rates are significantly higher than those published by most laboratories so far (7,21,22).…”

“…The first birth of a rhesus macaque after in vitro fertilization (IVF) was reported in 1984 (7), but the development of ARTs in nonhuman primates has been slow compared with its human counterpart. The efficacy of IVF methods is critically dependent on the protocols used for ovarian stimulation, in vitro embryo culture, embryo transfer, and uterine-embryo synchrony.…”

Chromosomal instability in rhesus macaque preimplantation embryos

“…Procedures for oocyte recovery, sperm collection, insemination and embryo culture have been described previously (22)(23)(24). Briefly, follicular fluid aspirates were collected in TALP-Hepes containing 0.3% bovine serum albumin (BSA).…”

In Vitro Manipulation of Nonhuman Primate Gametes for Embryo Production and Embryo Transfer.

“…Establishing an efficient ET technique is therefore an important task in this research field. In 1984, 6 years after the first successful ET in man [94], ET in nonhuman primates was successful in the rhesus monkey [7], the baboon [17] and the cynomolgus monkey [5]. My research group also used ET to the oviduct of cynomolgus monkeys and successfully generated offspring (unpublished data) (Fig.…”