Screening of conditions for rapid freezing of human oocytes: preliminary study toward their cryopreservation

One hundred and twenty-one freshly-collected human oocytes and 839 unfertilized human oocytes after insemination were cryopreserved by vitrification. The cryoprotectants used were dimethylsulfoxide (DMSO) and sucrose. Vital staining and morphological criteria were used to assess injuries to cells. Variation of the time exposure to DMSO and sucrose, and cryoprotectants concentrations, followed by extraction-dilution in sucrose without freezing made it possible to study chemical toxicities. Variation of cryoprotectant concentrations followed by immersion in liquid nitrogen, thawing, extraction, and dilution made it possible to choose optimal conditions for vitrification. The sucrose concentration upon extraction after freezing and thawing which was lower than that during soaking enhanced the oocyte survival rate as did the choice of duration and temperature of soaking. No parthenogenetical activation of these unfertilized ovum was observed. This study indicates that with a certain combination of DMSO and sucrose concentrations up to 80% of morphologically intact human oocytes can be recovered after rapid freezing and thawing.