THE IN-VITRO DEVELOPMENT OF MOUSE EMBRYOS BEYOND THE BLASTOCYST STAGEINTO THE HATCHING AND OUTGROWTH STAGE USING DIFFERENT ENERGY-SOURCES

Purpose: The purpose of this study was to investigate the effect of ma le and female serum supplementation on the in vitro development of mou se embryos beyond the blastocyst stage until the outgrowth stage since the latter may be related to the nidation of the embryo. We also stud ied the effect of EGF addition on embryo culture and blastocyst outgro wth. Methods and Results: The blastocyst and hatching rates of two-cel l 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 eith er 50% BSA or 50% female serum (P < 0.01 and P < 0.05, respectively. I n the last experiment, either 100 or 150 ng/ml EGF was added to the cu lture medium from the two-cell stage till blastocyst development and t he latter were cultured till outgrowth in 508 BSA, male serum or femal e serum. For both concentrations of EGF: the outgrowth rate was signif icantly higher in male serum compared to the other conditions (P < 0.0 1 and P < 0.05, respectively. The outgrowth rate was also higher when EGF was used compared to plain medium before transferring the blastocy sts to either male of female serum (P < 0.01 for both). Conclusions: W e conclude that the development of embryos to the outgrowth stage is s ignificantly 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.