In this study, in vitro preimplantation embryo culture media especially for outbred stock mice (Institute of Cancer Research (ICR)) were optimized with different concentrations of Ethylenediaminetetraacetic acid (EDTA). A plot with embryo development rates against EDTA concentrations ranging from 0 to 500µM showed a unique pattern with two characteristic peaks. 200µM was adopted as an optimal concentration of EDTA. The optimized media were also evaluated with two culture systems; conventional large volume culture system (1ml) and micro-droplet culture system. In the conventional large volume culture system, the blastocyst development rates were compared among three different media (F-10, KSOM and KSOM with the optimized 200µM EDTA). The rates were 0.4%, 16.7% and 57.6%, respectively. The development rates for micro-droplet (10µl) culture system was 73.9%. In conclusion, 200µM EDTA concentration in 10µl droplets in the KSOM medium was found as most suitable culture conditions for ICR mouse embryos as the blastocyst development rate was higher in micro-droplet culture system than in the traditional conventional large volume culture system. 

1. Different microdroplet culture volume: This spreadsheet includes the ICR mouse development rate (%) in different microdroplet culture volumes with KSOM and EDTA optimized KSOM media.
2. Large volume culture system: This spreadsheet includes the ICR mouse development rate (%) in different culture media and different EDTA concentrations in large volume culture system (1ml).
3. Microdroplet culture system: This spreadsheet includes the development rate (%) in KSOM and different EDTA concentrations in microdroplet culture system (10 microlitre).