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Successful cryopreservation of mouse blastocysts using a new vitrification solution
C A Valdez1, O Abas Mazni, Y Takahashi
1Department of Theriogenology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Journal of Reproduction and Fertility
|November 1, 1992
Summary
This study developed a new, effective vitrification solution for cryopreserving mouse blastocysts. The solution, containing ethylene glycol, dimethyl sulfoxide, and 1,3-butanediol, achieved high survival and development rates, with no significant difference from controls after transfer to recipients.
Area of Science:
- Reproductive Biology
- Cryobiology
- Developmental Biology
Background:
- Cryopreservation of mammalian embryos is crucial for assisted reproductive technologies.
- Cryoprotective agents (CPAs) are essential for preventing ice crystal formation during cryopreservation.
- Toxicity and efficacy of different CPAs and their combinations require thorough investigation for optimal embryo cryopreservation.
Purpose of the Study:
- To evaluate the toxicity of various permeating cryoprotectants on mouse blastocysts.
- To develop and validate a novel, effective vitrification solution for mouse blastocyst cryopreservation.
- To assess the developmental potential of cryopreserved mouse blastocysts post-warming.
Main Methods:
- Mouse blastocysts were exposed to six different permeating cryoprotectants at various concentrations and durations.
- Toxicity assays were performed to identify the least harmful cryoprotectant.
- A new vitrification solution (VSv) composed of ethylene glycol, dimethyl sulfoxide, and 1,3-butanediol was formulated and tested.
- Embryos were cryopreserved using one-step or two-step equilibration protocols and vitrified.
- Post-warming survival and in vitro development rates were assessed.
- Vitrified-warmed embryos were transferred to pseudopregnant recipients to evaluate in vivo development.
Main Results:
- Ethylene glycol exhibited the least toxicity among the tested cryoprotectants.
- The novel vitrification solution (VSv) demonstrated high in vitro developmental rates (95.4-96.2%) with optimized exposure times.
- Cryopreservation using VSv resulted in normal fetal development rates comparable to non-vitrified controls (45.0-54.2% vs. 60.0%).
- Prolonged exposure to cryoprotectant solutions negatively impacted embryo development.
Conclusions:
- A simple and effective vitrification solution (VSv) was successfully developed for mouse blastocyst cryopreservation.
- The developed VSv, utilizing ethylene glycol, dimethyl sulfoxide, and 1,3-butanediol, offers a promising alternative for embryo cryopreservation.
- Optimized exposure times are critical for maximizing the success of blastocyst cryopreservation using this vitrification method.