Identification of storage conditions stabilizing extracellular vesicles pre
File(s)
Author(s)
Type
Journal Article
Abstract
Extracellular vesicles (EVs) play a key role in many physiological and pathophysiological processes and hold great potential for therapeutic and diagnostic use. Despite significant advances within the last decade, the key issue of EV storage stability remains unresolved and under investigated. Here, we aimed to identify storage conditions stabilizing EVs and comprehensively compared the impact of various storage buffer formulations at different temperatures on EVs derived from different cellular sources for up to 2 years. EV features including concentration, diameter, surface protein profile and nucleic acid contents were assessed by complementary methods, and engineered EVs containing fluorophores or functionalized surface proteins were utilized to compare cellular uptake and ligand binding. We show that storing EVs in PBS over time leads to drastically reduced recovery particularly for pure EV samples at all temperatures tested, starting already within days. We further report that using PBS as diluent was found to result in severely reduced EV recovery rates already within minutes. Several of the tested new buffer conditions largely prevented the observed effects, the lead candidate being PBS supplemented with human albumin and trehalose (PBS-HAT). We report that PBS-HAT buffer facilitates clearly improved short-term and long-term EV preservation for samples stored at -80°C, stability throughout several freeze-thaw cycles, and drastically improved EV recovery when using a diluent for EV samples for downstream applications.
Date Issued
2022-06-01
Date Acceptance
2022-05-16
ISSN
2001-3078
Publisher
Co-Action Publishing
Journal / Book Title
Journal of Extracellular Vesicles
Volume
11
Issue
6
Copyright Statement
© 2022 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.
This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/35716060
Subjects
diluent
exosomes
extracellular vesicles
liposomes
preservation
stability
storage
storage buffer
vesicles
Extracellular Vesicles
Freezing
Humans
Nucleic Acids
Trehalose
Humans
Trehalose
Nucleic Acids
Freezing
Extracellular Vesicles
diluent
exosomes
extracellular vesicles
liposomes
preservation
stability
storage
storage buffer
vesicles
Extracellular Vesicles
Freezing
Humans
Nucleic Acids
Trehalose
0601 Biochemistry and Cell Biology
Publication Status
Published
Coverage Spatial
United States
OA Location
https://onlinelibrary.wiley.com/doi/10.1002/jev2.12238
Article Number
ARTN e12238
Date Publish Online
2022-06-18