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Human Reproduction, Vol 12, 1259-1262, Copyright © 1997 by Oxford University Press

ARTICLES

The optimal equilibration time for mouse embryos frozen by vitrification with trehalose

GM Ishida, H Saito, N Ohta, T Takahashi, MM Ito, T Saito, K Nakahara and M Hiroi
Department of Obstetrics and Gynecology, Yamagata University School of Medicine, Iida-Nishi, Japan.

To determine the best equilibration time in the cryoprotectant before rapid cooling, 8-cell mouse embryos were exposed to a vitrification solution containing ethylene glycol, Ficoll and trehalose in modified phosphate-buffered saline at 5 degrees C for varying periods of time. They were frozen using an ultra rapid freezing method, thawed in a 20 degrees C water bath and cultured for 24 h with 5-bromo-2-deoxyuridine. Embryo development and the number of sister chromatid exchanges, a sensitive indicator of genetic damage, were observed. The results demonstrated that embryo development after freezing and thawing was similar among the groups exposed for periods of 5-40 min. However, the number of sister chromatid exchanges was significantly smaller in the group exposed for 5 min, indicating that this was the safest equilibration time in the vitrification solution.
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