Effects of various low temperatures, cryoprotective agents and cooling rates on the survival, fertilizability and development of frozen-thawed mouse eggs

Abstract

Frozen mouse eggs were examined to determine the effects of low temperatures, concentration of cryoprotective agents and cooling rates on their survival, fertilizability in vitro and subsequent development. Dimethyl sulfoxide administered at 1.5 M concentration was found to be the most effective cryoprotective agent. Upon thawing, 51% and 56% of the eggs appeared to be normal after having been cooled at 0.33 degrees C/min to -30 degrees C and -50 degrees C, but only 18% of the eggs appeared to be normal after having been cooled at the same rate to -75 degrees C. When eggs were cooled at 0.33 degrees C/min to -45 degrees C and the cooling rate increased to 1 degree C/min from -45 degrees C to -75 degrees C, 44% and 72% appeared normal upon thawing. Of the normal eggs fertilized in vitro from C3H mice, 65% cleaved to the 2-cell stage and 24% of the 2-cell eggs developed into blastocysts. Following the transfer of 17 blastocysts into three recipient mice, one mouse delivered three normal young.