A quick pipeline for the isolation of 3D cell culture‐derived extracellular vesicles

Recent advances in cell biology research regarding extracellular vesicles have highlighted an increasing demand to obtain 3D cell culture‐derived EVs, because they are considered to more accurately represent EVs obtained in vivo. However, there is still a grave need for efficient and tunable methodo...

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Published inJournal of extracellular vesicles Vol. 11; no. 10; pp. e12273 - n/a
Main Authors Kyykallio, Heikki, Faria, Alessandra V. S., Hartmann, Rosabella, Capra, Janne, Rilla, Kirsi, Siljander, Pia R‐M
Format Journal Article
LanguageEnglish
Published United States John Wiley & Sons, Inc 01.10.2022
John Wiley and Sons Inc
Wiley
Subjects
3D cell culture
Breast cancer
cancer spheroids
Cell culture
Cell Culture Techniques, Three Dimensional
Cellulose
Experiments
Extracellular Vesicles
Humans
Hydrogels
isolation
Melanoma
nanofibrillar cellulose
Neoplasms
Penicillin
Polyethylene glycol
Spheroids
Technical Note
Italy
United States > US
Switzerland
Germany
Finland
3D cell culture
isolation
extracellular vesicles
cancer spheroids
nanofibrillar cellulose
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Summary:Recent advances in cell biology research regarding extracellular vesicles have highlighted an increasing demand to obtain 3D cell culture‐derived EVs, because they are considered to more accurately represent EVs obtained in vivo. However, there is still a grave need for efficient and tunable methodologies to isolate EVs from 3D cell cultures. Using nanofibrillar cellulose (NFC) scaffold as a 3D cell culture matrix, we developed a pipeline of two different approaches for EV isolation from cancer spheroids. A batch method was created for delivering high EV yield at the end of the culture period, and a harvesting method was created to enable time‐dependent collection of EVs to combine EV profiling with spheroid development. Both these methods were easy to set up, quick to perform, and they provided a high EV yield. When compared to scaffold‐free 3D spheroid cultures on ultra‐low affinity plates, the NFC method resulted in similar EV production/cell, but the NFC method was scalable and easier to perform resulting in high EV yields. In summary, we introduce here an NFC‐based, innovative pipeline for acquiring EVs from 3D cancer spheroids, which can be tailored to support the needs of variable EV research objectives.
Bibliography:Heikki Kyykallio and Alessandra V. S. Faria contributed equally to this study.
Kirsi Rilla and Pia R.‐M. Siljander contributed equally to this study.
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ISSN:2001-3078
2001-3078
DOI:10.1002/jev2.12273