Normal Calves Obtained after Direct Transfer of Vitrified Bovine Embryos Using Ethylene Glycol, Trehalose, and Polyvinylpyrrolidone

Abstract

In the present study, IVF bovine embryos were vitrified using as the cryoprotectants, ethylene glycol plus trehalose plus the polymer, polyvinylpyrrolidone (PVP). In Experiment I, toxicity of the vitrification solution (VS) containing 20% PVP was tested in relation to temperature and exposure time. One hundred percent embryo development was observed with treatment at 57C for 5 min, whereas only 55.5% embryos were developed when the treatment was carried out at 207C for 5 min. In Experiment II, embryos were vitrified using one of the three treatments (Treatment A, 40% ethylene glycol (EG); treatment B, 40% EG / 11.3% trehalose; and treatment C, 40% EG / 11.3% trehalose / 20% PVP and rehydrations) was performed directly in mPBS. Highest development (84.1%) and hatching rate (68.2%) were obtained when embryos were vitrified with the vitrification solution used in treatment C. In Experiment III, embryos were vitrified as in Experiment II (treatment C). The development and hatching rates were compared after rehydration in different rehydration solutions. No significant difference was observed among the development and hatching rates when rehydration was carried out in different concentrations of trehalose. Five vitrified–warmed bovine embryos were transferred directly to five recipients and three recipients gave birth to three normal calves.