Embryo cryopreservation in cynomolgus monkeys.

An improved knowledge of cryopreservation of primate embryos will have important research and clinical application. Fifty-six 4- to 8-cell in vitro fertilized embryos were frozen in HEPES-buffered Tyrode's solution containing 1.5 M dimethylsulfoxide (DMSO) and cooled at the rate of 0.3 degrees C/minute to -39 degrees C before being transferred into liquid nitrogen. Embryos were rapidly thawed at room temperature for 2 minutes. DMSO was diluted with medium in three steps at 5-minute intervals. Of the 56 embryos, 39 (70%) were classified as viable on the basis of surviving the freezing process with greater than 50% of their blastomeres intact. Twelve of the 39 embryos were cultured overnight, and 11 cleaved at least once. Twenty-five embryos were transferred to nine synchronized, unstimulated recipient monkeys 24 to 48 hours after ovulation. Three pregnancies (33.3%) resulted from the nine transfers.