Abstract
Purpose
To clarify the developmental capacity of frozen two-cell blastomeres, we investigated in vivo and in vitro viabilities of blastomeres that were frozen ultrarapidly after separation from two-cell mouse embryos. Two-cell embryos obtained from superovulated F1 hybrid females were denuded by treatment with 0.5% pronase solution and then induced to separate into two single blastomeres by gentle pipetting. The blastomeres were cryopreserved by an ultrarapid freezing method.
Results
The preimplantation developmental rate of two-cell embryos frozen in 3.0 MDMSO was significantly higher than the rate of those frozen in 15 and 4.5 MDMSO (at least P<0.05). The in vitro developmental rate of the ultrarapidly frozen-thawed blastomeres separated from two-cell embryos (75.0%) was similar to that of nonfrozen blastomeres (76.0%). When eight pairs of blastocysts that developed from frozen two-cell mouse blastomeres were transferred to pregnant ICR recipients on Day 3, four live singletons were born.
Conclusion
Thus, the results indicate that two-cell mouse blastomeres can be frozen by the ultrarapid freezing method.
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Kang, MJ., Han, YM., Lee, CS. et al. Cryopreservation of blastomeres separated from two-cell mouse embryos by an ultrarapid freezing method. J Assist Reprod Genet 11, 409–413 (1994). https://doi.org/10.1007/BF02211728
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DOI: https://doi.org/10.1007/BF02211728