Search

Purpose: The purpose of this study was to investigate the effect of male and female serum supplementation on the in vitro development of mouse embryos beyond the blastocyst stage until the outgrowth stage since the latter may be related to the nidation of the embryo. We also studied the effect of EGF addition on embryo culture and blastocyst outgrowth. Methods and Results: The blastocyst and hatching rates of two-cell mouse embryos cultured in Ham’s F-10 + BSA, Ham’s F-10 + male serum, or Ham’s F-10 + female serum were found to be comparable (P > 0.05). The outgrowth rate of hatched blastocysts was significantly increased, though, when they were transferred to 50% male serum compared to either 50% BSA or 50% female serum (P < 0.01 and P < 0.05, respectively. In the last experiment, either 100 or 150 ng/ml EGF was added to the culture medium from the two-cell stage till blastocyst development and the latter were cultured till outgrowth in 508 BSA, male serum or female serum. For both concentrations of EGF: the outgrowth rate was significantly higher in male serum compared to the other conditions (P < 0.01 and P < 0.05, respectively. The outgrowth rate was also higher when EGF was used compared to plain medium before transferring the blastocysts to either male of female serum (P < 0.01 for both). Conclusions: We conclude that the development of embryos to the outgrowth stage is significantly enhanced by male serum. The addition of EGF from the two-cell stage also significantly improves the outgrowth success rate for bath male and female serum conditions.

The role of growth hormone (GH) in follicular development, ovulation and embryo development is currently under reconsideration. In this study, we have tried to investigate the effect of GH on preimplantation development. of mouse embryos in vitro. Zygotes and two-cell mouse embryos were cultured without (control) or with GH. For zygotes, the addition of 0.2 mu g/ml of GH resulted in 77 +/- 1% of blastocysts formed and 66 +/- 3% rate of hatching (control 64 +/- 4 and 31 +/- 3%, p < 0.05 and p < 0.01, respectively). For two-cell embryos, the addition of 0.2 mu g/ml of GH resulted in 87 +/- 2% of blastocysts formed and 60 +/- 4% hatching rate (control 76 +/- 4 and 47 +/- 5%, p < 0.05 for both). This positive effect of GH addition implies that the latter can support mouse preimplantation development in vitro and it suggests, along with its local action on the ovary and its possible effects, via the insulin-like growth factor system, on the tubal and uterine epithelium, a continuous role of this hormone in reproductive physiology from follicular maturation to embryonic development and, possibly, implantation.

Purpose: This study deals with the suitability of Ham’s F-10 without hypoxanthine for early cleavage-stage embryo culture. A high percentage of mouse two-cell embryos developed into morula and blastocysts in Ham’s F-10 formulated without hypoxanthine (75.3 and 71.6%, respectively); in contrast, in agreement with previous reports, only 15.4% developed beyond the two-cell stage in Ham’s F-10 with hypoxanthine. To begin to evaluate the effect of hypoxanthine on human embryos, a total of 318 human oocytes was fertilized and cultured in Ham’s F-10 minus hypoxanthine. Results: The fertilization, cleavage, and pregnancy percentages in two ovulation induction protocols [human menopausal gonadotropin (hMG)/human chorionic gonadotropin (hCG) and gonadotropin releasing hormone/hMG/hCG] were 82, 88, and 20 and 83, 91, and 30%, respectively. Conclusion: These results suggest that, in agreement with mouse embryo development, hypoxanthine does not appear to be necessary for human embryo cleavage and its omission from Ham’s F-10 may enhance the conditions for culture of early-cleaving human embryos. These observations may lead to a better understanding of critical cell processes during early human embryonic development.

During a 12-year period, a variety of fetal diagnostic techniques were performed in 112 twin, 3 triplet and 1 quintuplet pregnancies, respectively. Tissues sampled included amniotic fluid, fetal blood and chorionic villi. Two spontaneous abortions occurred and one case of twins was stillborn. Four selective feticides were performed in twins for specific indications and four healthy surviving infants were delivered. It is concluded that fetal diagnosis in multiple pregnancy is safe and accurate without significant perinatal morbidity and mortality.