Experiments were conducted to find a suitable cryoprotectant and suitable procedure for vitrification of 8-cell mouse embryos. The method was then applied clinically to the cryopreservation of human embryos in our assisted reproduction programme. Mouse embryos were vitrified with 30 or 40% 1,2-propanediol (PROH), dimethylsulphoxide (DMSO), ethylene glycol, glycerol, or acetamide, each diluted with a solution containing 30% Ficoll plus 0.5 M sucrose. Embryos were exposed to the solutions for 0.5 or 2 min at 20 or 25°C, cooled in liquid nitrogen and warmed rapidly. Embryo survival was assessed by in-vitro development. In PROH-, DMSO- and acetamide-based solutions, higher survival rates (29–82%) were obtained with less permeating conditions, suggesting that these cryoprotectants are considerably toxic. In glycerol-and ethylene glycol-based solutions, however, higher survival rates (74 and 92% respectively) were obtained with more permeating conditions, suggesting that these cryoprotectants are less toxic. Human embryos on days 2–3 were vitrified in an ethylene glycol-based solution (EFS40). Survival, assessed by the morphology, was higher in 4-cell embryos on day 2 and 8-cell embryos on day 3 than in 2–3-cell embryos on day 2 or 2–7-cell embryos on day 3. From 18 transfers, one ended with the delivery of healthy twin babies.
Copyright (C) 1998, European Society of Human Reproduction and Embryology
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