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Systematic review of factors influencing extracellular vesicle yield from cell cultures

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Abstract

The potential therapeutic utility of extracellular vesicles (EVs) has spawned an interest into a scalable production, where the quantity and purity of EV samples is sufficient for clinical applications. EVs can be isolated using several different protocols; however, these isolation protocols and the subsequent methods of quantifying the resulting EV yield have not been sufficiently standardized. Therefore, the possibility of comparing different studies with respect to these parameters is limited. In this review, we have presented factors that might influence the yield and function of EVs from cell culture supernatants. The methods of isolation, downstream quantification, and culture conditions of the EV producing cells have been discussed. In order to examine the inter-study coherency of EV yields, 259 studies were initially screened, and 46 studies were included for extensive downstream analysis of EV yields where information pertaining to the isolation protocols and quantification methods was obtained from each study. Several other factors influencing yield were compared, such as cell type producing EVs, cell confluence level, and cell stimulation. In conclusion, various factors may impact the resulting EV yield, including technical aspects such as EV isolation and quantification procedures, and biological aspects such as cell type and culture conditions. The reflections presented in this review might aid in future standardization of the workflow in EV research.

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Abbreviations

EV:

Extracellular vesicle

MVB:

Multivesicular body

UC:

Ultracentrifugation

SEC:

Size-exclusion chromatography

UC-W:

Ultracentrifugation with washing step

UC-G:

Ultracentrifugation with gradient separation

FCS:

Fetal calf serum

CM:

Conditioned medium

BCA:

Bicinchoninic acid

NTA:

Nanoparticle tracking analysis

FA:

Fixed angle

SW:

Swinging bucket

DLS:

Dynamic light scattering

TRPS:

Tunable resistive pulse sensing

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  1. Laboratory of Cancer Biology, Institute of Health Science and Technology, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg Ø, Denmark

    Johann Mar Gudbergsson, Martin Najbjerg Skov & Meg Duroux

  2. Laboratory of Neurobiology, Institute of Health Science and Technology, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg Ø, Denmark

    Kasper Bendix Johnsen

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  1. Johann Mar Gudbergsson
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Gudbergsson, J.M., Johnsen, K.B., Skov, M.N. et al. Systematic review of factors influencing extracellular vesicle yield from cell cultures. Cytotechnology 68, 579–592 (2016). https://doi.org/10.1007/s10616-015-9913-6

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