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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9579059/ https://www.ncbi.nlm.nih.gov/pubmed/36257915 http://dx.doi.org/10.1002/jev2.12273 |
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author | Kyykallio, Heikki Faria, Alessandra V. S. Hartmann, Rosabella Capra, Janne Rilla, Kirsi Siljander, Pia R‐M |
author_facet | Kyykallio, Heikki Faria, Alessandra V. S. Hartmann, Rosabella Capra, Janne Rilla, Kirsi Siljander, Pia R‐M |
author_sort | Kyykallio, Heikki |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9579059 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-95790592022-10-19 A quick pipeline for the isolation of 3D cell culture‐derived extracellular vesicles Kyykallio, Heikki Faria, Alessandra V. S. Hartmann, Rosabella Capra, Janne Rilla, Kirsi Siljander, Pia R‐M J Extracell Vesicles Technical Note 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. John Wiley and Sons Inc. 2022-10-18 2022-10 /pmc/articles/PMC9579059/ /pubmed/36257915 http://dx.doi.org/10.1002/jev2.12273 Text en © 2022 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles. https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Technical Note Kyykallio, Heikki Faria, Alessandra V. S. Hartmann, Rosabella Capra, Janne Rilla, Kirsi Siljander, Pia R‐M A quick pipeline for the isolation of 3D cell culture‐derived extracellular vesicles |
title | A quick pipeline for the isolation of 3D cell culture‐derived extracellular vesicles |
title_full | A quick pipeline for the isolation of 3D cell culture‐derived extracellular vesicles |
title_fullStr | A quick pipeline for the isolation of 3D cell culture‐derived extracellular vesicles |
title_full_unstemmed | A quick pipeline for the isolation of 3D cell culture‐derived extracellular vesicles |
title_short | A quick pipeline for the isolation of 3D cell culture‐derived extracellular vesicles |
title_sort | quick pipeline for the isolation of 3d cell culture‐derived extracellular vesicles |
topic | Technical Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9579059/ https://www.ncbi.nlm.nih.gov/pubmed/36257915 http://dx.doi.org/10.1002/jev2.12273 |
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