Journal
CELLULAR AND MOLECULAR LIFE SCIENCES
Volume 76, Issue 12, Pages 2369-2382Publisher
SPRINGER BASEL AG
DOI: 10.1007/s00018-019-03071-y
Keywords
Exosomes; Purification; Isolation methods; Nanomedicine; Theranostics
Categories
Funding
- Fundacio La Marato de TV3 [201516-10, 201502-30]
- SGR programme of Generalitat de Catalunya [2017-SGR-301, 2017-SGR-483]
- ISCIII-REDinREN [RD16/0009]
- Instituto Carlos III [PI17/00336]
- Instituto de Salud Carlos III [PI17/01487, PIC18/00014]
- Red de Terapia Celular-TerCel [RD16/00111/0006]
- CIBER Cardiovascular projects, Plan Nacional de I+D+I [CB16/11/00403]
- ISCIII-Subdireccion General de Evaluacion y el Fondo Europeo de Desarrollo Regional (FEDER)
- PERIS from the Generalitat de Catalunya [SLT002/16/00234]
- la Caixa Banking Foundation
- Societat Catalana de Cardiologia
- ACCI (Catalonia Trade Investment
- Generalitat de Catalunya) under the Catalonian ERDF (European Regional Development Fund) operational program 2014-2020
- Health Department of the Catalan Government (Generalitat de Catalunya)
- Spanish Ministry of Economy and Competitiveness-MINECO [SAF2017-84324-C2-1-R]
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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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