The dependence of blastocyst development on the extracellular Na+/K+ ratio was investigated in an in vitro system. Hatching and outgrowth of mouse blastocysts was enhanced at Na+/K+ ratios between 3 and 10 compared to the ratio of about 25 typical for most culture media and serum. At a Na+/K+ ratio of 2, blastocyst hatching and outgrowth were inhibited. The requirement of blastocyst development for relatively high extracellular K+ concentrations agrees with the fact that K+ concentrations in oviduct and uterine secretions are higher than in serum. The findings can also be relevant in optimizing in vitro culturing techniques for blastocysts.