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Ovarian steroid replacement therapy in the ovariectomized ewe, given in the correct sequence to mimic endogenous steroid changes in the normal ovulatory cycle, allows the development of embryos transferred in utero. A similar type of sequential therapy was designed for steroid replacement in women with primary ovarian failure. This produces the histological changes in uterine endometrial morphology and plasma oestradiol and progesterone similar to those observed in the normal ovulatory cycle. We now report that in one of these women a donated oocyte, fertilized by her husband's spermatozoa and cultured to the two-cell stage in vitro, was transferred in utero, resulting in a normal pregnancy and the delivery of a healthy child. Oestrogen therapy was withdrawn at 12 weeks and progesterone at 19 weeks gestation. This technique allows the treatment of human infertility due to primary ovarian failure.

Summary. Oocytes were obtained from patients with tubal infertility at fixed times after the onset of the endogenous LH rise or hCG injection, and were inseminated immediately after recovery or after periods of 4\ p=n-\ 4\ m=1/ 2\ , 5\ p=n-\ 5\ m=1/ 2\ and 6\ p=n-\ 6\ m=1/ 2\ h in culture in vitro. Delayed insemination resulted in a marked increase in the proportion of oocytes that were fertilized and developed to normal embryos and maximum rates occurred after 5\ p=n-\ 5\ m=1/ 2\ h in culture ( 0\ p=n-\ \ m=1/ 2\ h, 26%; 4\ p=n-\ 4\ m=1/ 2\ h, 50%; 5\ p=n-\ 5\ m=1/ 2\ h, 89%; 6\ p=n-\ 6\ m=1/ 2\ h, 69%). The range and mean (\m=+-\s.d.) intervals from insemination for the pronuclear and early cleavage stages were 27\p=n-\43(35\m=.\6\m=+-\4\m=.\4)h for 2-cell stages, 36\p=n-\65 (45\m=.\7\ m=+-\ 8\m=.\3) h for 4-cell stages, 45\p=n-\73 (54\m=.\3\ m=+-\ 12\m=.\6) h for 8-cell stages and 68\p=n-\85 h for the 16-cell stage. In 7/50 patients receiving 1 or 2 embryos at the 2-, 4-and 8-cell stages, fetal development was normal and 2 women had twin pregnancies (36% success compared with 8% for single embryos). All pregnancies were from the groups in which insemination was delayed for 5\ p=n-\ 6\ m=1/ 2\ h. It is concluded that a short period of culture in vitro may allow the completion of oocyte maturation, and improve the results of in-vitro fertilization.

In vitro fertilisation after stimulation of the ovulatory cycle has led to successful pregnancy. If more oocytes are recovered than are needed they may be left unfertilised, preserved, or donated to a recipient couple from whom oocytes cannot be obtained. A case of human pregnancy initiated by transfer of a donated embryo fertilised in vitro is reported. The donor was a 42 year old woman with primary infertility from whom six follicles were aspirated after stimulation of the ovulatory cycle. The recipient was a 38 year old infertile woman who had undergone several unsuccessful attempts for artificial insemination from a donor. Five oocytes were recovered from the donor's six follicles, four of which were inseminated with spermatozoa of the donor's husband and the fifth with a frozen sample of semen. Three of the four embryos fertilised by her husband were returned to the donor and the fifth was transferred to the recipient. No pregnancy was recorded in the donor, but pregnancy was confirmed in the recipient, though spontaneous abortion occurred after 10 weeks. This case will give useful information for further study of in vitro fertilisation, but also raises many ethical issues.

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