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

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.

Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

“…All types of cells cultured in vitro release EVs and other factors into their culture medium, thus creating cell culture‐conditioned medium [CCM; Shekari et al., 2023]. This includes eukaryotic cells from multi‐ and unicellular organisms and prokaryotic cells including gram‐positive and ‐negative bacteria and Mycobacteria .…”

Inter‐laboratory multiplex bead‐based surface protein profiling of MSC‐derived EV preparations identifies MSC‐EV surface marker signatures

“…The dynamic relationship between culture medium components and EV production remains a focal point in EV research, shaping its course toward clinical translation (Shekari et al., 2023 ). Notably, the culture medium is enriched with essential supplements such as fetal bovine serum, human serum and human platelet lysate, which serve as both critical growth factors and prominent sources of EVs and EV‐like particles.…”

Systems Approaches to Cell Culture-Derived Extracellular Vesicles for Acute Kidney Injury Therapy: Prospects and Challenges

“… 31–33 On the other hand, cells in culture can be characterized, validated, and grown under defined, xeno-free conditions for EV generation, thus allowing for greater consistency. 34 , 35 Furthermore, scientific and legislative frameworks for cell therapy are well established and multiple human clinical trials of MSC therapies have already been completed in AKI and other diseases. As such, this review article focuses mostly on EVs derived from the culture of human MSCs, as we believe these have the highest likelihood of translation to clinical use.…”

“… 41 Due to the effects of such variables, recommendations for reporting experimental parameters have been released by the ISEV, which can be consulted for more information. 35 …”