Abstract
The method of vitrification has been widely used for cryopreservation. However, the effectiveness of this method for mammalian oocytes could be improved by optimizing each step of the process. In the present study, we tested the effects of varying several key parameters to determine the most effective protocol for mouse oocyte vitrification. We found that cryoprotectant containing ethylene glycol and dimethylsulfoxide plus 20% fetal calf serum produced the highest rates of oocyte survival, fertilization, and blastocyst formation. The duration and temperature of oocyte exposure to vitrification and thawing solutions influenced survival rate. The presence of cumulus cells surrounding oocytes and the incubation of thawed oocytes in Toyoda-Yokoyama-Hosoki medium also increased oocyte survival. Open pulled straw and nylon loop methods were more effective than the mini-drop method. Finally, the combination of these improved methods resulted in better spindle morphology when compared to the unimproved methods. These results demonstrate that the outcomes of mouse oocyte vitrification can be improved by a suitable combination of cryopreservation methods, which could be applied to future clinical research with human oocytes.
Highlights
Gamete cryopreservation has become the leading technology for long-term germplasm preservation
High concentrations of cryoprotectant inside cells become hyaloid material under ultra-cooling conditions, which prevents the formation of ice crystals that are harmful to cells
A high concentration of cryoprotectant has toxic effects that may lead to oocyte damage
Summary
Gamete cryopreservation has become the leading technology for long-term germplasm preservation. Oocytes in the control group showed greater survival (92.1 ± 3.6%), fertilization (91.2 ± 5.5%), and blastocyst formation (74.2 ± 5.9%) than all other groups. Effect of concentration of FCS on oocyte survival, fertilization, and blastocyst formation.
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