Vitrification for bovine embryos with low-quality grade

This study was conducted to examine the utility of vitrification for bovine embryos with low-quality grade, and simple cryoprotectants dilution method for practitioners. In Experiment 1, survival of frozen embryos was compared with that of vitrified embryos using minimum volume cooling (MVC). Then, vitrified embryos were used to confirm the optimum sucrose concentration in Experiment 2. The survival rates of embryos that had been vitrified following diluted cryoprotectants with the one-step in-straw method were compared with those of fresh control embryos in Experiment 3. Frozen-thawed or vitrified-warmed blastocysts were cultured with TCM-199 supplemented with 100 μmol/L beta-mercaptoethanol +5% fetal bovine serum at 38.5°C in an atmosphere of 5% CO₂ in air, their survival after 24 hr were compared. The development to term of fair quality in vivo embryos after vitrification was examined in Experiment 4. Results show that survival rates of frozen-thawed embryos were lower (p < .05) than that of vitrified-warmed ones. When vitrified embryos were warmed in 0.3 mol/L sucrose in straws, their survival rate was 100%. The total cell numbers of vitrified-warmed embryos were comparable to those of fresh control embryos. The six calves from 13 vitrified embryos were delivered in Experiment 4. These results indicate that MVC vitrification following one-step cryoprotectants dilution is utilized to preserve low-quality bovine embryos.