Effect of cryopreservation technique and season on the survival of in vitro produced cattle embryos

Embryo cryopreservation is a major tool for conservation and propagation of genetically superior animals. However, it adversely affects the survival of embryos. The objective of this study was to determine the effects of cryopreservation technique (vitrification compared with slow freezing) and different seasons in which oocytes were obtained on the post-warming survival of in vitro produced (IVP) cattle morulae. In experiment 1, morulae (Day 6 post-IVF), obtained from abattoir-sourced oocytes during spring, summer, fall and winter over a period of 3.5 years, were subjected to either vitrification (n=271 morulae), slow freezing (n=281 morulae) or no freezing (control; n=249 morulae). After warming, the morulae were cultured to the expanded blastocyst stage (Day 8 post-IVF). Data were compared using Glimmix procedure in SAS®. Blastocyst rate differed (P<0.05) among the treatments: unfrozen control (78±3.6%), vitrification (52±4.6%) and slow freezing (35±4.2%). The re-expansion of vitrified morulae upon warming was not correlated with subsequent blastocyst rate (r=-0.048; P>0.05). The morulae produced during fall season had lesser (P<0.05) cleavage and morula rates (67±1.6%; Day 2 post-IVF and 22±1.4%; Day 6 post-IVF, respectively) than all other seasons (74±1.1 and 30±1.2%, respectively). Blastocyst rate was the least (P<0.05) when oocytes were collected during the summer season in both control and slowly frozen groups. Blastocyst development rate did not change due to season in vitrification group (P>0.05). In conclusion, vitrification is a more desirable technique than slow freezing for cryopreservation of IVP cattle morulae. If the slow freezing method is employed, greater success can be achieved using oocytes collected in the winter and spring with a primary contributing factor being lesser morulae development if oocytes are collected in the fall and also the lesser blastocyst formation of cryopreserved morulae when oocytes are collected in the summer.