Does storage time in LN2 influence survival and pregnancy outcome of vitrified rabbit embryos?

Vitrification is one of the most widely used techniques for embryo cryopreservation. The aim of this work was to study the effect of storage time in liquid nitrogen on vitrified rabbit embryos. A total of 1467 vitrified rabbit embryos were transferred into 174 females. The embryos had been maintained in liquid nitrogen during 3 different periods, A) < 1 year (98 transfers, 827 embryos); B) 2-5 years (44 transfers, 360 embryos) and C) > 15 years (32 transfers, 280 embryos). A generalized linear model was used to determine the effect of period on pregnancy and birth rates. A Bayesian approach was applied to analyze the survival rate of the vitrified embryos. In all analyses the number of transferred embryos was included as covariate. It was observed that neither the period of storage nor the number of transferred embryos affected pregnancy rate, and all periods presented similar pregnancy rates (0.85 ± 0.04; 0.86 ± 0.05; 0.78 ± 0.07 for A, B and C). Fertility at birth was affected by the number of transferred embryos, but non-significant differences between periods were detected (0.77 ± 0.04, 0.75 ± 0.07; 0.69 ± 0.08 for A, B and C). Also, the posterior means (highest posterior density intervals at 95%) of embryo survival at birth from pregnant females were similar between the different periods (47 [41 53]; 47 [38 56]; 42 [31 54]; for A, B and C). Results obtained in the present experiment point out that vitrified embryos could be stored in liquid nitrogen during at least fifteen years, achieving good pregnancy rate, fertility and survival at birth.