Abstract
The following three methods were evaluated in order to obtain a most efficient freezing protocol for the preservation of two-cell mouse embryos: (a) slow cooling and slow thawing in 1.5M dimethyl sulfoxide, (b) slow cooling and fast thawing in 1.5M propanediol (PROH), and (c) ultrarapid freezing and fast thawing in either 3.5M DMSO or 3.0M PROH. In the slow-cooling procedures (a and b) ice nucleation (seeding) was induced manually or automatically. With method a, only a slight difference, 51.8% for manual and 58.9% for automatic seeding, was observed in survival rates, while the development to blastocysts was significantly affected: 35.4% with manual and less than 10% with automatic induction (P<0.001). Method b gave high survival (86.2%) and developmental rates (69.0%) with manual seeding compared with automatic seeding (20.7 and 9.8%, respectively;P<0.001). Using protocol c, higher survival and developmental rates were obtained with DMSO (84.8 and 55.9%) than with PROH (39.8 and 19.4%,P<0.001). These results demonstrate that inducing nucleation manually is superior to the use of a highly sophisticated autoseeding system and that method b with manual seeding is most effective in preserving the developmental capacity of twocell mouse embryos after freezing and thawing. There is evidence that this is also true of human embryo cryopreservation.
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Macas, E., Xie, M., Keller, P.J. et al. Developmental capacities of two-cell mouse embryos frozen by three methods. J Assist Reprod Genet 8, 208–212 (1991). https://doi.org/10.1007/BF01130806
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DOI: https://doi.org/10.1007/BF01130806