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Preliminary results are presented concerning the first clinical application of cocultures of human embryos. In the experimental group, the embryos were cultured and transferred on day 5 post-insemination. Blastocyst formation was not dependent upon the stimulation regimen. Long term or ultrashort stimulation of the ovaries after gonadotrophin releasing hormone analogue gave 55-60% blastocyst formation. Serum was not necessary to obtain blastocysts. When the embryos were cocultured, we observed an increase in the implantation rates per embryo in the pregnant patients. However, a real increase in the pregnancy rate per transfer was observed in a population of patients who had had repeated failures of embryo transfer. This observation is discussed as possibly bypassing an effect of uterine motility, but the overall beneficial effect has to be assessed in a double blind randomized study. It is probable that improvements will not be observed for all the indications for in vitro fertilization.

This study proposes a procedure for the isolation and culture of oviduct epithelial cells of several species. In-vitro culture on such a feeder seems to allow full embryonic development and viability. The inner linings of Fallopian tubes from mouse, rabbit, cow and human were trypsinized and the epithelial cells were enriched with Percoll gradient. Isolated cells, obtained in high yield with good viability, were maintained in monolayer culture in B2-Menezo medium supplemented with serum, which also supports early embryonic development in vitro. The plated primary cultures reached confluence within 8 days, producing a monolayer of cohesive polygonal cells. Associated with this large epithelial cell population, ciliated cells as well as polykaryotic cells and few fibroblastic nests were observed. After the first sub-culture, the ciliated cells disappeared and the epithelial cell monolayer grew rapidly to confluence within 3 days and displayed contact inhibition. No epithelial cell growth could be obtained in culture in the absence of serum. The addition of oestrogens had no effect on any of the cultured oviductal epithelial cells. A spontaneous alteration was observed in morphology and growth after several passages, the number of which depends mainly upon the species.

Two progesterone presentations, a vaginal application of 90 mg progesterone per day (Crinone) or 300 mg progesterone administered orally (Utrogestan), were compared for luteal phase support of patients undergoing an in-vitro fertilization (IVF) procedure. A total of 283 patients were randomly allocated to either treatment. The treatment started within 24 h after the embryo transfer procedure and continued until day 30 in cases of implantation. Efficacy was assessed using the pregnancy and delivery rates. Safety was assessed through specific symptoms and usual safety monitoring. The pregnancy rates per transfer were not significantly different in the Crinone and Utrogestan groups at days 12 (Crinone 35.3%, Utrogestan 29.9%, P = 0.55), 30 (Crinone 28.5%, Utrogestan 25.0%, P = 0.61) and 90 (Crinone 25.9%, Utrogestan 22.9%, P = 0.69). No differences in the spontaneous abortion rates were seen thereafter. The delivery rates (number of deliveries per patient; Crinone 23.0%, Utrogestan 22.2%, P = 1.00), as well as the ratio of newborn babies per embryo transferred (Crinone 11.7%, Utrogestan 11.1%, P = 0.91), were not significantly different. Safety parameters were similar in both groups, except for drowsiness, which was more significantly frequent in the oral progesterone group than in the Crinone group at all time points. No serious adverse events were recorded in this study. The fact that Crinone matches the efficacy of the larger doses of progesterone used orally reflects an advantage of the transvaginal route of administration which avoids the metabolic inactivation of progesterone during its first liver pass.

Purpose : To determine if hypergonadotropic hypogonadism related to galactosemia could be linked to anomaly of the circulating FSH. A 26-year-old woman, suffering GALT (Galactoso-1-phosphate uridyltransferase) had a premature ovarian failure with amenorrhea since the age of 19. The circulating level for FSH was 83 and 34 mU/mL for LH. Methods : After treatment with a hormonal substitution cycle including estradiol and progesterone, the patient underwent stimulations with recombinant FSH. The first cycle, one 16-mm diameter follicle and the second cycle one follicle of 17.5 mm of diameter were obtained at the time of ovulation induction. Results : The patient conceived and delivered a female baby weighting 3.38 kg after the second stimulation protocol. Conclusions : The impact of galactosemia on the ovary seems rather related to the absence of recognition of circulating FSH by its receptor and not to a toxic alteration of the ovary by itself as it is currently reported. The rFSH treatment following hormonal substitution cycles allows to overcome infertility problems.

Purpose:To determine if GV oocytes, collected at the time of ICSI, can be matured in vitro and rescued for therapeutic treatment. A patient for whom all the collected oocytes at the GV stage after a classical COH protocol were matured in vitro with GH. Method: All the naked oocytes were matured in a culture medium (ISM2) containing 15% patient serum +1.6 units of GH (Saizen) per millilitre. Oocytes were incubated overnight at 37 • C. The MII oocytes obtained were micro-injected. A fresh transfer was performed and a supernumerary blastocyst was frozen. Results: The patient was pregnant and delivered a healthy girl after transfer of the frozen/thawed blastocyst. The baby girl is now 2 years old. Conclusion: In vitro maturation with GH allows rescuing naked GV oocytes collected at the time of ICSI. GH action does not pass through the cumulus cells. According to the possible lack of synchrony between the embryo and the uterus, we recommend to freeze the embryos obtained and to replace them in a controlled cycle.

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