The effect on fertilization of exposure of mouse oocytes to dimethyl sulfoxide: an optimal protocol

Mouse oocytes, with or without an intact cumulus mass, were exposed to various concentrations of dimethyl sulfoxide (DMSO) at different temperatures for different periods of time and using different protocols of DMSO addition and removal. The effect of these procedures on the chymotrypsin sensitivity of the zona pellucida and the fertilizability of the oocytes was then assessed. Some procedures were found to affect adversely both the zona pellucida and the cumulus mass, resulting in reductions in the fertilization rate. As a result of both these and previously reported experiments (1-3), an optimal schedule is proposed for the handling of mouse oocytes during cryopreservation, namely, to equilibrate cumulus-intact oocytes in 1.5 M DMSO precooled to 4 degrees C prior to freezing, to remove DMSO at 4 degrees C after thawing prior to restoring the oocytes to 37 degrees C, to loosen or remove the cumulus cells, and then to hold oocytes at 37 degrees C for at least 1 hr to allow recovery of the spindle prior to insemination.