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. 2023 Oct 16;2(10):e115.
doi: 10.1002/jex2.115. eCollection 2023 Oct.

Cell culture-derived extracellular vesicles: Considerations for reporting cell culturing parameters

Faezeh Shekari  1   2 , Faisal J Alibhai  3 , Hossein Baharvand  1   4 , Verena Börger  5 , Stefania Bruno  6 , Owen Davies  7 , Bernd Giebel  5 , Mario Gimona  8 , Ghasem Hosseini Salekdeh  9 , Lorena Martin-Jaular  10 , Suresh Mathivanan  11 , Inge Nelissen  12 , Esther Nolte-'t Hoen  13 , Lorraine O'Driscoll  14 , Francesca Perut  15 , Stefano Pluchino  16 , Gabriella Pocsfalvi  17 , Carlos Salomon  18 , Carolina Soekmadji  19 , Simon Staubach  20 , Ana Claudia Torrecilhas  21 , Ganesh Vilas Shelke  22 , Tobias Tertel  5 , Dandan Zhu  23 , Clotilde Théry  10 , Kenneth Witwer  24 , Rienk Nieuwland  25   26
Affiliations

Cell culture-derived extracellular vesicles: Considerations for reporting cell culturing parameters

Faezeh Shekari et al. J Extracell Biol. .

Abstract

Cell culture-conditioned medium (CCM) is a valuable source of extracellular vesicles (EVs) for basic scientific, therapeutic and diagnostic applications. Cell culturing parameters affect the biochemical composition, release and possibly the function of CCM-derived EVs (CCM-EV). The CCM-EV task force of the Rigor and Standardization Subcommittee of the International Society for Extracellular Vesicles aims to identify relevant cell culturing parameters, describe their effects based on current knowledge, recommend reporting parameters and identify outstanding questions. While some recommendations are valid for all cell types, cell-specific recommendations may need to be established for non-mammalian sources, such as bacteria, yeast and plant cells. Current progress towards these goals is summarized in this perspective paper, along with a checklist to facilitate transparent reporting of cell culturing parameters to improve the reproducibility of CCM-EV research.

Keywords: cell culture‐conditioned medium; cellular therapy; ectosomes; exosomes; extracellular vesicles; in vitro; reproducibility; rigor; standardization.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
Source and application of extracellular vesicles. (a) Extracellular vesicles (EVs) can be prepared from biological fluids, cell culture‐conditioned medium (CCM), or ex vivo tissue. (b) Following molecular and morphological characterization, EVs can be used to study intercellular communication biology, to diagnose diseases (biomarker research), or as therapeutic agents (directly or as drug delivery vehicles). The dotted box indicates the focus of the CCM‐derived EV (CCM‐EV) task force on cell culturing parameters and their effects on EV release, content, and biological function.
FIGURE 2
FIGURE 2
Cell culture parameters and their effects on production of extracellular vesicles. (a) All parameters affect cells during conditioning and release of extracellular vesicles (EVs). (b) Parameters influencing release of EVs, and (c) biochemical composition of EVs. (d) Cells might also re‐package cell culture medium supplement components into/onto released EVs. (e) Cell culturing parameters may affect processes that indirectly affect EVs.
See this image and copyright information in PMC

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