Abstract
Extracellular vesicles (EV) are small membrane structures released by cells that act as potent mediators of intercellular communication. The study of EV biology is important, not only to strengthen our knowledge of their physiological roles, but also to better understand their involvement in several diseases. In the field of biomedicine they have been studied as a novel source of biomarkers and drug delivery vehicles. The most commonly used method for EV enrichment in crude pellet involves serial centrifugation and ultracentrifugation. Recently, different protocols and techniques have been developed to isolate EV that imply less time and greater purification. Here we carry out a comparative analysis of three methods to enrich EV from plasma of healthy controls: ultracentrifugation, ExoQuickTM precipitation solution (System Biosciences), and Total Exosome Isolation kit (Invitrogen). Our results show that commercial precipitation reagents are more efficient and enable higher EV enrichment factors compared with traditional ultracentrifugation, although subsequent imaging analysis is not possible with some of them. We hope that this work will contribute to the current research on isolation techniques to assist the progress of clinical applications with diagnostic or therapeutic objectives.
Highlights
Extracellular vesicles (EV) are small membrane-bound particles containing proteins, lipids and nucleic acids from donor cells that may be functional in recipient cells.Recently EV have emerged as means of communication between distant cells
Different types of EV or EV from different sources require different enrichment or isolation approaches depending on the downstream studies
The free protein contamination in the case of biological fluids must be taken into account
Summary
EV have emerged as means of communication between distant cells They are produced by several mechanisms: by fusion of multivesicular bodies and the plasma membrane (usually referred as exosomes), or directly by plasma membrane budding in response to intracellular or extracellular stimuli [1,2,3]. A proper and standardized method of isolation and characterization of EV is required to deepen our biological understanding of EV and their potential use as biomarkers [4], or to explore their possibilities as drug delivery systems [5,6]. The source of EV (biological fluids or cell culture supernatants), the starting volumes, or the possibility to scale up or down according to the number of vesicles required for further analysis are, among others, factors to consider when choosing an EV isolation protocol. From the variety of methods to isolate EV currently available (reviewed by Li et al [11]), the choice of the most suitable one, taking into account subsequent EV characterization and analysis, is crucial
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
