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Trehalose delivered by cold-responsive nanoparticles improves tolerance of cumulus-oocyte complexes to microwave drying

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Abstract

Purpose

Trehalose is a non-permeable protectant that is the key to preserve live cells in a dry state for potential storage at ambient temperatures. After intracellular trehalose delivery via cold-responsive nanoparticles (CRNPs), the objective was to characterize the tolerance of cat cumulus-oocyte complexes (COCs) to different levels of microwave-assisted dehydration.

Methods

Trehalose was first encapsulated in CRNPs. After exposure to trehalose-laden CRNPs, different water amounts were removed from cat COCs by microwave drying. After each dehydration level, meiotic and developmental competences were evaluated via in vitro maturation, fertilization, and embryo culture. In addition, expressions of critical genes were assessed by quantitative RT-PCR.

Results

CRNPs effectively transported trehalose into COCs within 4 h of co-incubation at 38.5 °C followed by a cold-triggered release at 4 °C for 15 min. Intracellular presence of trehalose enabled the maintenance of developmental competence (formation of blastocysts) as well as normal gene expression levels of HSP70 and DNMT1 at dehydration levels reaching up to 63% of water loss.

Conclusion

Intracellular trehalose delivery through CRNPs improves dehydration tolerance of COCs, which opens new options for oocyte storage and fertility preservation at ambient temperatures.

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Data availability

Data are available upon request to the corresponding author.

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Acknowledgements

We thank Dr. Keiko Antoku (Last Chance Animal Rescue) and Dr. Joy Lewis (Spay Now Animal Surgery Clinic), and their staff for providing domestic cat testes and ovaries.

Funding

This work was supported by the Office of the Director, National Institutes of Health, grant/award number: R01OD023139. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Authors and Affiliations

Authors

Contributions

PL: conceptualization, project administration, methodology, investigation, formal analysis, visualization, and writing—original draft preparation; SS: methodology and writing—methodology; OA: investigation, analysis, writing—methodology, and review; HS: methodology and writing—review; XH: resources and writing—review; PC: conceptualization, funding acquisition, resources, supervision, and writing—review and editing.

Corresponding author

Correspondence to Pierre Comizzoli.

Ethics declarations

Ethical disclosure

The Animal Care and Use Committee from the Smithsonian’s National Zoo and Conservation Biology Institute granted a waiver of the animal care and use approval for that study because testes and ovaries were collected at local veterinary clinics as byproducts from owner-requested routine neutering and spaying.

Conflict of interest

The authors declare no competing interests.

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Supplementary Information

Suppl. Fig. 1

High resolution image (TIF 12966 kb)

10815_2023_2831_MOESM1_ESM.tif

Kinetics of water content during microwave drying. A 40 µl drop of 0.3 M trehalose was microwave dried for 0 to 40 min (5 min interval). Water contents were expressed as gH2O/gDW (left Y axis) and percentage of water weight (right Y axis). Values are mean ± SEM. (PNG 87 kb)

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Lee, PC., Stewart, S., Amelkina, O. et al. Trehalose delivered by cold-responsive nanoparticles improves tolerance of cumulus-oocyte complexes to microwave drying. J Assist Reprod Genet 40, 1817–1828 (2023). https://doi.org/10.1007/s10815-023-02831-x

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  • DOI: https://doi.org/10.1007/s10815-023-02831-x

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