Cryopreservation of mouse 2-cell embryos and ova by vitrification: Methodologic studies

S. Friedler, E. Shen, E. J. Lamb

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Cryopreservation of unfertilized mouse ova and 2-cell embryos by a vitrification technique was examined. Survival was defined by development to the hatching blastocyst stage after in vitro fertilization. With 19 embryos at the 2-cell stage, the authors obtained 100% morphologic survival and 89% development to hatching blastocyst stage. To define the optimal conditions for vitrification of ova, the authors treated a total of 845 unfertilized ova. In experiments done at 0°C, the concentration of vitrification solution (VS1) and the length of exposure of ova to VS1 both had significant (P < 0.01) effects on survival. The mean survival rate for controls in ten experiments was 52%. VS1 100% or 90% in HEPES buffered saline and 10 minutes' exposure yielded rates that did not differ significantly from controls. Significantly lower survival rates followed the use of 70 and 80% solution and exposure for 5, 15, 20, or 30 minutes. Thus, under these conditions, exposure of unfertilized mouse ova to VS1 and cooling to 0°C did not interfere with in vitro fertilization and development of embryos. However, in five experiments in which a total of 101 ova were plunged into liquid nitrogen after treatment with VS1 under the optimal conditions, none could be fertilized in vitro.

Original languageEnglish
Pages (from-to)306-314
Number of pages9
JournalFertility and Sterility
Issue number2
StatePublished - 1 Jan 1987
Externally publishedYes


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