Cryopreservation of bovine oocytes: is cryoloop vitrification the future to preserving the female gamete?

Abstract

The cryoloop is a technique where a thin nylon loop is used to suspend a film of cryoprotectant containing the oocytes and directly immersing them in liquid nitrogen. 508 bovine oocytes were collected, of these 351 were cryopreserved by slow freezing using standard straws or a new vitrification method using our self-constructed cryoloops and the remainder were controls. After thawing, the oocytes were inseminated by ICSI or standard IVF. The cryoloop vitrification method yielded a survival rate of 90.5% and the slow freezing technique a rate of 54.4% (p < 0.0001). When ICSI was performed, cryopreservation by the cryoloop vitrification method resulted in very similar cleavage rate to controls (16.0% vs. 17.3%) but slow freezing produced a slightly lower rate (9.4%). Cleavage rates after IVF in fresh oocytes was higher than the cryopreservation groups (49.5% vs. 15.4% and 25.8%), whereas after ICSI the rates were similar in all groups (17.3% vs. 9.4% and 16%). It is concluded that the new cryoloop vitrification technique followed by ICSI produce good embryo formation results and they could hold the future for effective oocyte cryopreservation.