Comparison of traditional and modified (VitMaster) methods of rabbit embryo vitrification

Abstract

In spite of their cryobiological efficacy, minimum-volume vitrification methods suffer from the risk of microbiological contamination and are technically and/or manually demanding. In this study, the effects of a traditional, slightly modified vitrification method and vitrification using supercooled liquid nitrogen (VitMaster) applied for rabbit morula-stage embryos were compared. Embryos were equilibrated in a solution containing 1,2-propanediol (2.72 M) and glycerol (1.36 M) for 7 min and vitrified in 0.25-ml insemination straws after 1-min exposure to a vitrification solution containing additionally 1.0 M sucrose. Cooling was performed in 'normal' or supercooled liquid nitrogen. Regardless of the cooling method applied, high in vitro survival and development rates of vitrified embryos were obtained. All embryos were intact after warming, and 61 out of 65 (93.8%) and 23 out of 24 (95.8%) embryos developed to the blastocyst stage after 48-h in vitro culture of embryos vitrified in 'normal' or supercooled liquid nitrogen, respectively. The results suggest higher developmental ability of embryos vitrified in supercooled liquid nitrogen (91.7% vs . 83.1% of embryos vitrified traditionally developed to more advanced, expanding and/or hatching blastocyst stages). In vivo survival rate, tested for the traditional vitrification system only, revealed that 36.8% of embryos developed to term. The results show promise for establishing a fully successful method for rabbit embryo vitrification.