Identification of storage conditions stabilizing extracellular vesicles preparations
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 investigat...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022-06-01
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| Series: | Journal of Extracellular Vesicles |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jev2.12238 |
| _version_ | 1811241277074702336 |
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| author | André Görgens Giulia Corso Daniel W. Hagey Rim Jawad Wiklander Manuela O. Gustafsson Ulrika Felldin Yi Lee R. Beklem Bostancioglu Helena Sork Xiuming Liang Wenyi Zheng Dara K. Mohammad Simonides I. van de Wakker Pieter Vader Antje M. Zickler Doste R. Mamand Li Ma Margaret N. Holme Molly M. Stevens Oscar P. B. Wiklander Samir EL Andaloussi |
| author_facet | André Görgens Giulia Corso Daniel W. Hagey Rim Jawad Wiklander Manuela O. Gustafsson Ulrika Felldin Yi Lee R. Beklem Bostancioglu Helena Sork Xiuming Liang Wenyi Zheng Dara K. Mohammad Simonides I. van de Wakker Pieter Vader Antje M. Zickler Doste R. Mamand Li Ma Margaret N. Holme Molly M. Stevens Oscar P. B. Wiklander Samir EL Andaloussi |
| author_sort | André Görgens |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T13:34:08Z |
| format | Article |
| id | doaj.art-b0afdd0ac68d4599b414eeb023f4d490 |
| institution | Directory Open Access Journal |
| issn | 2001-3078 |
| language | English |
| last_indexed | 2024-04-12T13:34:08Z |
| publishDate | 2022-06-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Vesicles |
| spelling | doaj.art-b0afdd0ac68d4599b414eeb023f4d4902022-12-22T03:31:05ZengWileyJournal of Extracellular Vesicles2001-30782022-06-01116n/an/a10.1002/jev2.12238Identification of storage conditions stabilizing extracellular vesicles preparationsAndré Görgens0Giulia Corso1Daniel W. Hagey2Rim Jawad Wiklander3Manuela O. Gustafsson4Ulrika Felldin5Yi Lee6R. Beklem Bostancioglu7Helena Sork8Xiuming Liang9Wenyi Zheng10Dara K. Mohammad11Simonides I. van de Wakker12Pieter Vader13Antje M. Zickler14Doste R. Mamand15Li Ma16Margaret N. Holme17Molly M. Stevens18Oscar P. B. Wiklander19Samir EL Andaloussi20Department of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Cardiology Experimental Cardiology Laboratory University Medical Center Utrecht, Utrecht University Utrecht The NetherlandsDepartment of Cardiology Experimental Cardiology Laboratory University Medical Center Utrecht, Utrecht University Utrecht The NetherlandsDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Medical Biochemistry and Biophysics Karolinska Institutet Stockholm SwedenDepartment of Medical Biochemistry and Biophysics Karolinska Institutet Stockholm SwedenDepartment of Medical Biochemistry and Biophysics Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Clinical Research Center Karolinska Institutet Stockholm SwedenAbstract 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.https://doi.org/10.1002/jev2.12238diluentexosomesextracellular vesiclesliposomespreservationstability |
| spellingShingle | André Görgens Giulia Corso Daniel W. Hagey Rim Jawad Wiklander Manuela O. Gustafsson Ulrika Felldin Yi Lee R. Beklem Bostancioglu Helena Sork Xiuming Liang Wenyi Zheng Dara K. Mohammad Simonides I. van de Wakker Pieter Vader Antje M. Zickler Doste R. Mamand Li Ma Margaret N. Holme Molly M. Stevens Oscar P. B. Wiklander Samir EL Andaloussi Identification of storage conditions stabilizing extracellular vesicles preparations Journal of Extracellular Vesicles diluent exosomes extracellular vesicles liposomes preservation stability |
| title | Identification of storage conditions stabilizing extracellular vesicles preparations |
| title_full | Identification of storage conditions stabilizing extracellular vesicles preparations |
| title_fullStr | Identification of storage conditions stabilizing extracellular vesicles preparations |
| title_full_unstemmed | Identification of storage conditions stabilizing extracellular vesicles preparations |
| title_short | Identification of storage conditions stabilizing extracellular vesicles preparations |
| title_sort | identification of storage conditions stabilizing extracellular vesicles preparations |
| topic | diluent exosomes extracellular vesicles liposomes preservation stability |
| url | https://doi.org/10.1002/jev2.12238 |
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