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Survival rate of Mus domesticus domesticus blastocysts vitrified in 9.0m of ethylene glycol with sucrose

The aim of these experiments was to determine the in vitro and in vivo survivability of Mus domesticus domesticus blastocysts, following exposure and vitrification in modified PBS solution containing 9.0M of ethylene glycol (EG) and 0.3M of sucrose. The cryoprotectant solutions were denominated I (without sucrose) and IS (with sucrose) when the embryo exposure to the solutions has been done in two steps. First the embryos were exposed to a modified PBS solution with 1,8M EG + 6% BSA during 2 minutes, and after they were transferred to straws containing modified PBS plus 9.0M EG + 6% BSA and after 30 seconds immersed into liquid nitrogen. The others cryoprotectant solutions, denominated II (without sucrose) and IIS (with sucrose) the embryos were exposed directly to the 9.0M EG and after 30 seconds plunged into the liquid nitrogen. In the first experiment a survival rate of 299 blastocysts was observed following exposure to cryoprotectant solutions. There was no statistical diference among control and treatment groups. The experiment II allowed to evaluate the in vitro embryo survival, after vitrification of 330 blastocysts, the cryoprotectant solutions I and IS determinated statistically different (p<0.05) embryo survival rates, showing in vitro hatching rates of 49 and 40%, respectively. In experiment 3, 141 blastocysts were exposed to the cryoprotectant solutions I and IS, vitrified and then transferred into the recipients. The number of implantations and fetuses were observed after 14 days of gestation. There was no statistical difference (p>0.05) among the solutions I and IS in the number of implantations (37 and 32%) and fetuses (27 and 27%), respectively. The survival rate of Mus domesticus domesticus blastocysts did not improve with the addition of sucrose to the vitrification solution containing 9.0M of EG.

rodent; embryo; cryopreservation


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