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Extracellular vesicle isolation methods: rising impact of size-exclusion chromatography

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Abstract

Extracellular vesicles (EVs) include a variety of nanosized vesicles released to the extracellular microenvironment by the vast majority of cells transferring bioactive lipids, proteins, mRNA, miRNA or non-coding RNA, as means of intercellular communication. Remarkably, among other fields of research, their use has become promising for immunomodulation, tissue repair and as source for novel disease-specific molecular signatures or biomarkers. However, a major challenge is to define accurate, reliable and easily implemented techniques for EV isolation due to their nanoscale size and high heterogeneity. In this context, differential ultracentrifugation (dUC) has been the most widely used laboratory methodology, but alternative procedures have emerged to allow purer EV preparations with easy implementation. Here, we present and discuss the most used of the different EV isolation methods, focusing on the increasing impact of size exclusion chromatography (SEC) on the resulting EV preparations from in vitro cultured cells-conditioned medium and biological fluids. Comparatively, low protein content and cryo-electron microscopy analysis show that SEC removes most of the overabundant soluble plasma proteins, which are not discarded using dUC or precipitating agents, while being more user friendly and less time-consuming than gradient-based EV isolation. Also, SEC highly maintains the major EVs’ characteristics, including vesicular structure and content, which guarantee forthcoming applications. In sum, together with scaling-up possibilities to increase EV recovery and manufacturing following high-quality standards, SEC could be easily adapted to most laboratories to assist EV-associated biomarker discovery and to deliver innovative cell-free immunomodulatory and pro-regenerative therapies.

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Data source: EV-TRACK database, August 2018 [89]

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Acknowledgements

This work was supported in part by Fundació La Marató de TV3 (201516-10, 201502-30), SGR programme of Generalitat de Catalunya (2017-SGR-301 REMAR Group, and 2017-SGR-483 ICREC Group), ISCIII-REDinREN (RD16/0009 Feder Funds) and Instituto Carlos III (PI17/00336), grants from the Spanish Ministry of Economy and Competitiveness—MINECO (SAF2017-84324-C2-1-R), Instituto de Salud Carlos III (PI17/01487, PIC18/00014), Red de Terapia Celular-TerCel (RD16/00111/0006), CIBER Cardiovascular (CB16/11/00403) projects, as part of the Plan Nacional de I + D+I, and co-funded by ISCIII-Subdirección General de Evaluación y el Fondo Europeo de Desarrollo Regional (FEDER). MMT is sponsored by the PERIS (SLT002/16/00234) from the Generalitat de Catalunya, “la Caixa” Banking Foundation, and Societat Catalana de Cardiologia. This work has been developed in the context of AdvanceCat with the support of ACCIÓ (Catalonia Trade & Investment; Generalitat de Catalunya) under the Catalonian ERDF (European Regional Development Fund) operational program 2014–2020; FEB is a researcher from Fundació Institut de Recerca en Ciències de la Salut Germans Trias i Pujol, supported by the Health Department of the Catalan Government (Generalitat de Catalunya).

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Authors and Affiliations

  1. REMAR-IVECAT Group, Germans Trias i Pujol Health Science Research Institute, Can Ruti Campus, Badalona, Spain

    Marta Monguió-Tortajada & Francesc E. Borràs

  2. ICREC Research Program, Germans Trias i Pujol Health Science Research Institute, Can Ruti Campus, Badalona, Spain

    Marta Monguió-Tortajada, Carolina Gálvez-Montón, Antoni Bayes-Genis & Santiago Roura

  3. Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Bellaterra, Spain

    Marta Monguió-Tortajada & Francesc E. Borràs

  4. CIBERCV, Instituto de Salud Carlos III, Madrid, Spain

    Carolina Gálvez-Montón, Antoni Bayes-Genis & Santiago Roura

  5. Cardiology Service, Germans Trias i Pujol University Hospital, Badalona, Spain

    Antoni Bayes-Genis

  6. Department of Medicine, UAB, Barcelona, Spain

    Antoni Bayes-Genis

  7. Nephrology Service, Germans Trias i Pujol University Hospital, Badalona, Spain

    Francesc E. Borràs

Authors
  1. Marta Monguió-Tortajada
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  2. Carolina Gálvez-Montón
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  3. Antoni Bayes-Genis
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  4. Santiago Roura
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  5. Francesc E. Borràs
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Contributions

MM-T, SR and FB wrote the manuscript; MM-T, CG-M, AB-G, SR and FB: critical reading and final approval of the manuscript. All authors reviewed the manuscript.

Corresponding authors

Correspondence to Santiago Roura or Francesc E. Borràs.

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Monguió-Tortajada, M., Gálvez-Montón, C., Bayes-Genis, A. et al. Extracellular vesicle isolation methods: rising impact of size-exclusion chromatography. Cell. Mol. Life Sci. 76, 2369–2382 (2019). https://doi.org/10.1007/s00018-019-03071-y

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  • DOI: https://doi.org/10.1007/s00018-019-03071-y

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