Survival rates of mouse blastocyst vitrified in dimethylformamide based solutions associated with ethylene glicol or 1-2 propanediol

Abstract

The aim of this study was to determine the effect of dimethylformamide (DF) associated with ethylene glycol (EG) or 1-2 propanediol (PROH) during vitrification, on the in vitro development of mouse blastocysts. Cryoprotectant toxicity was evaluated exposing embryos into three different equilibrium solutions (ES) composed by DF, EG or PROH mixtures (10% v/v of each) in mPBS + 0.5% PVA at different interval times (1, 3 and 10min). In a second experiment, embryos were exposed to the same ES (either 1 or 3min), following for the three respectively vitrification solutions (VS) (20% v/v of each) for 30s. After 72 hours of in vitro culture, embryo hatching and expansion rates were similar for the ES1 and ES2 equilibration solutions during the time interval of 1 or 3min. However embryos exposed for 10 min to the DF equilibration solutions, had lower survival rates than EG-PROH solution (P<0.01). Furthermore, survival rates for embryos exposed to DF-PROH (ES+VS) were lower than embryos exposed to the other solutions (P<0.01). Blastocyst vitrification was performed with the three ES+VS (for 1min and 30s, respectively), using glass micropipettes (GMP). Survival rates were lower for blastocysts vitrified with DF solutions (3%-3/108 and 17.1%-19/111) (P<0.01) than with PROH+EG vitrification solutions (69%-73/105). In conclusion, DF as a cryoprotectant into vitrification solutions have deleterious effects on the in vitro developmental competence of vitrified mouse blastocysts.

Survival and developmental competence of bovine embryos at different developmental stages and separated blastomeres after vitrification in different solutions

“…These results support those obtained by Villmil et al (2011). Also, experiment 1 of the present study showed that, although the survival of the EG-vitrified blastocyst group was significantly lower than that of the group vitrified in the EG + PG combination, hatching ability did not significantly differ between the two groups.…”

“…Also, experiment 1 of the present study showed that, although the survival of the EG-vitrified blastocyst group was significantly lower than that of the group vitrified in the EG + PG combination, hatching ability did not significantly differ between the two groups. These results support those obtained by Villmil et al (2011). Moreover, the total number of cells in blastocysts vitrified with EG + PG was significantly higher than that of EG alone.…”

“…These results support those obtained by Villmil et al . (). Moreover, the total number of cells in blastocysts vitrified with EG + PG was significantly higher than that of EG alone.…”