Correlation between the cryosurvival, cell number and diameter in bovine in vitro produced embryos

Abstract

The selection of quality embryos is a prerequisite of cryopreservation process. Present study was conducted to examine the correlation between the diameter and cryotolerance, on the cell number of the cryopreserved embryos. The blastocyst stage embryos were collected at culture days 7–9, evaluated morphologically under a microscope and divided according to the diameter into three groups: Group 1; (larger than 150 μm), Group 2; (diameter of 100–150 μm), Group 3; (smaller than 100 μm). Blastocysts were vitrified-thawed using the classical vitrification method and then cultured in SOF medium drops at 24 h. Blastocysts were considered viable if they re-expanded or hatched from the zona pellucidae. Finally re-expanded blastocysts from the Group 1 and Group 2 to determine the differential count of cells in the ICM and TE. The re-expansion ability of blastocysts 100–150 μm in diameter (69.56%) was significantly higher than other groups (52.17 and 47.36%). The value of the correlation coefficient between the re-expansion rate and cell number of blastocysts in the group 2 (r = 0.784) tended to be higher than that in the group 1 (r = 0.512) and group 3 (r = 0.491) (p < 0.05). For ICM/total cell ratio yield group 2 embryos showed higher rate (0.28), compared to the other groups (0.19 and 0.16). In conclusion, the present study demonstrates that the correlation between diameter of embryos and their cryosurvival based on re-expansion ability and cell allocation.