Cryoprotection of Day-4 mouse embryos by methanol

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Abstract
Methanol was examined as a cryoprotective additive that permits the direct transfer of frozen-thawed day-4 mouse embryos to foster mothers without dilution of the cryoprotectant. Methanol permeated the embryos rapidly, was not toxic and exerted a cryoprotective action. The highest level of survival (50%) of embryos in vitro was observed after equilibration in Medium PB1 containing 3.0 M-methanol, slow cooling (0.5.degree. C/min) to a temperature between -30.degree. and -40.degree. C, rapid cooling (800.degree. C/min) and storage in liquid nitrogen (-196.degree. C), a rapid warming (800.degree. C/min), and rapid dilution. A high rate of development in vivo to late-stage fetuses (up to 81%) was observed when cryopreserved embryos were transferred to pseudopregnant recipients immediately after thawing.

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