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Effects of multi-gradient equilibration during vitrification on oocyte survival and embryo development in mice

Published online by Cambridge University Press:  24 November 2023

Yan Zhu ,
Zhen Zhang ,
Guang-Li Zhang  and
Man-Xi Jiang Open the ORCID record for Man-Xi Jiang [Opens in a new window]
Yan Zhu
Affiliation:
Medical Experimental Center, Guangdong Second Provincial General Hospital, Guangzhou 510317, People’s Republic of China
Zhen Zhang
Affiliation:
Medical Experimental Center, Guangdong Second Provincial General Hospital, Guangzhou 510317, People’s Republic of China
Guang-Li Zhang
Affiliation:
Center for Reproductive Medicine, Guangdong Second Provincial General Hospital, Guangzhou 510317, People’s Republic of China
Man-Xi Jiang*
Affiliation:
Center for Reproductive Medicine, Guangdong Second Provincial General Hospital, Guangzhou 510317, People’s Republic of China
*
Corresponding author: Man-Xi Jiang; Email: manxijiang@yahoo.com
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Summary

Vitrification has been widely used for oocyte cryopreservation, but there is still a need for optimization to improve clinical outcomes. In this study, we compared the routine droplet merge protocol with modified multi-gradient equilibration vitrification for cryopreservation of mouse oocytes at metaphase II. Subsequently, the oocytes were thawed and subjected to intracytoplasmic sperm injection (ICSI). Oocyte survival and spindle status were evaluated by morphology and immunofluorescence staining. Moreover, the fertilization rates and blastocyst development were examined in vitro. The results showed that multi-gradient equilibration vitrification outperformed droplet merge vitrification in terms of oocyte survival, spindle morphology, blastocyst formation, and embryo quality. In contrast, droplet merge vitrification exhibited decreasing survival rates, a reduced proportion of oocytes with normal spindle morphology, and lower blastocyst rates as the number of loaded oocytes increased. Notably, when more than six oocytes were loaded, reduced oocyte survival rates, abnormal oocyte spindle morphology, and poor embryo quality were observed. These findings highlight that the vitrification of mouse metaphase II oocytes by the modified multi-gradient equilibration vitrification has the advantage of maintaining oocyte survival, spindle morphology, and subsequent embryonic development.

Keywords

Embryo developmentFertilizationMulti-gradient equilibrationOocyte survivalOocyte vitrification
Type
Research Article
Information
Zygote , Volume 31 , Issue 6 , December 2023 , pp. 612 - 619
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Copyright
© The Author(s), 2023. Published by Cambridge University Press

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