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Vitrification of mammalian oocytes and embryos is typically a two-step procedure involving two solutions of increasing concentrations of cryoprotectants. In the present study, we report a simple vitrification protocol that uses low cryoprotectant concentration and a single medium (LCSM). This medium, along with the traditional high concentration two media (HCTM) protocol, was used to vitrify mouse oocytes, zygotes, and blastocysts using silica capillary, cryotop, cryolock, and 0.25 ml straws. Survival rates, two-cell rates, and blastocyst formation rates were compared for oocytes and zygotes vitrified using both protocols. Results show that the LCSM protocol was as good as or better than the traditional HCTM protocol for vitrifying mouse MII oocytes and zygotes using silica capillary, cryotop, and cryolock. On the other hand, for blastocysts, only silica capillary using LCSM had comparable results with the traditional HCTM protocol while cryolock and cryotop had significantly lower percentages of re-expanded and hatched blastocysts. Collapsing blastocysts prior to vitrification or longer duration for better cryoprotectant distribution in multicellular embryos may improve the outcome. In conclusion, the LCSM protocol, with one medium of much lower cryoprotectant concentrations and shorter equilibration time, reduces exposure to cryoprotectant toxicity while improves efficiency, consistency and reliability for mammalian oocyte and embryo preservation.
Bibliographical noteFunding Information:
We thank Dr. Charles Bormann at the ART Laboratory at Massachusetts General Hospital for providing Cryotop, Cryolock, and constructive criticism of the manuscript. This study was supported by funding from National Institutes of Health (grant number 5R24OD016985).
We thank Dr. Charles Bormann at the ART Laboratory at Massachusetts General Hospital for providing Cryotop, Cryolock, and constructive criticism of the manuscript. This study was supported by funding from National Institutes of Health (grant number 5R24OD016985 ).
© 2020 Elsevier Inc.
- Low cryoprotectant concentration
- Single medium
PubMed: MeSH publication types
- Journal Article
- Research Support, N.I.H., Extramural
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- 1 Active
ATP-Bio: NSF Engineering Research Center for Advanced Technologies for the Preservation of Biological Systems (ATP-Bio)
9/1/20 → 8/31/25
Project: Research project