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  • Title: Embryonic behavior of two-cell mouse embryos frozen by the one- and two-step ultrarapid techniques.
    Author: Vasuthevan S, Ng SC, Bongso A, Ratnam SS.
    Journal: J Assist Reprod Genet; 1992 Dec; 9(6):545-50. PubMed ID: 1299388.
    Abstract:
    PURPOSE: A modified two-step ultrarapid freezing technique was compared to the one-step ultrarapid freezing technique. Two-cell mouse embryos were frozen-thawed using the two freezing protocols, and postthaw cryoprotectant removal was carried out in either a single- or a multiple-step procedure. RESULTS: Statistically similar cryosurvival (96.95-100%) and blastocyst formation rates (87.95-91.47%) were obtained with both freezing groups. In addition, the method of cryoprotectant removal did not have any significant effect on the survival and development of the frozen-thawed embryos in both groups. Blastocysts formed following single-step cryoprotectant removal had significantly lower inner cell mass counts in the one-step than in the two-step group (26.14 and 27.59, respectively; P < 0.05). Embryo transfer studies showed that the implantation and fetal formation rates of embryos frozen by the two-step technique (61.67 and 60.0%, respectively) were similar to those of embryos frozen by the one-step technique (74.12 and 71.76%, respectively). CONCLUSION: These results demonstrate that the ultrarapid two-step technique is as effective in cryopreserving two-cell mouse embryos as the ultrarapid one-step technique.
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