Two-step freezing of two-cell rabbit embryos after partial dehydration at room temperature
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Summary. The effect of rapid freezing and thawing on the survival of 2-cell rabbit embryos was examined. When embryos in 2·2 m-propanediol were directly plunged from room temperature to liquid nitrogen some of them survived after thawing (8%) but only if they had been pretreated by exposure to an impermeable solute, sucrose, that makes the blastomeres shrink osmotically before cooling. High survival (77–88%) in vitro was obtained when pretreated embryos were first held at – 30°C for 30–240 min before immersion into liquid nitrogen. Transfer of such frozen—thawed embryos gave a survival rate to live young similar to that obtained with controls (26% and 32% respectively). DMSO was less effective than propanediol; only 2 out of 38 sucrose-pretreated frozen—thawed embryos developed in vitro.
The present work shows that a combination of partial dehydration of blastomeres at room temperature with their permeation by a cryoprotective agent offers a simple method for successful rapid freezing and thawing of rabbit embryos.
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