In an effort to improve the in vitro culture conditions of human gamates and embryos in our laboratory, this study was undertaken to assess the presence of hypoxanthine in Ham's Nutrient Mixture F-10, a chemically defined, complex medium,
commonly
used
for in vitro fertilization of human as well as animal oocytes, affects embryogenesis in our two-cell mouse embryo bioassay. For the two-cell mouse embryo bioassay, embryos extracted from each female mouse oviduct were equally distributed and
incubated
separately in Ham's F-10 with or without hypoxanthine supplementation, and either without any protein supplementation or with additional protein(4 mg/ml BSA or 10% human cord serum). Individual embryos were categorized as follows : 3-to 4-cell,
6-to
8-cell, morula, and blastocyst. Results were expressed as the proportion of embryos in each test dish achieving or surpassing the specified developmental stage at 24 hours intervals for a period of 72 hours.
@ES The results were as follows:
@EN 1. At 72 hours of incubation, 52.6% of the embryos without hypoxanthine reached the blastocyst stage, compared with 37.6%(p<0.05) for the Ham's F-10 with hypoxanthine. The proportion or embryos degenerating was significantly higher in the
culture
dishes containing hypoxanthine.
2. Development of 2-cell embryo to morula and blastocyst stage in vitro was not significantly different for the BSA supplementation in Ham's F-10 culture media, with or without hypoxanthine(p>0.05).
3. The addition of human cord serum supplementation to the Ham's F-10 without hypoxanthine was revealed a high percentage(73.1%) of the 2-cell embryos developed into blastocysts, compared with 58.9%(p<0.05) for the Ham's F-10 with hypoxanthine.
The conclusion of our results showed that Ham's F-10 medium containing hypoxanthine significantly reduced embryo development in the two-cell mouse embryo bioassay and the addition of human fetal cord serum increased the development of two cell
stage
mouse embryo to the blastocyst stage than the BSA supplementation.
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