Effects of various cryoprotectants on the survival of mouse embryos cryopreserved by the quick freezing method

The effects of concentrations of glycerol, ethylene glycol or dimethylsulphoxide (DMSO) in the presence of either 0.25 M lactose or sucrose on the post-thaw survival of mouse quickly-frozen compacted morulae were studied. In this method, the embryos were directly frozen in liquid nitrogen (LN2) vapor at approximately -170 degrees C for 2 min before being plunged into LN2. High survival rates of frozen-thawed embryos were obtained when the freezing medium contained 3 M ethylene glycol with either 0.25 M lactose or sucrose (76.5 and 70.2%, respectively). When the embryos were frozen in glycerol, significantly high survival was obtained with 3 M glycerol + 0.25 M sucrose (73.5%, P less than 0.001). However, a freezing medium containing DMSO with either sugar gave lower survival rates. At a higher concentration of 4 M, ethylene glycol with 0.25 M lactose gave significantly higher survival rate than glycerol or DMSO (P less than 0.05). Significantly higher rates were obtained at 2 M with all 3 cryoprotectants when the freezing medium contained lactose rather than sucrose (P less than 0.05). This study showed that glycerol and ethylene glycol were effective cryoprotectants in the quick freezing of mouse embryos, while DMSO was less effective. In addition, the protective effects of these cryoprotectants are affected by their concentrations and the type of sugar used.