Modern Techniques for the Isolation of Extracellular Vesicles and Viruses
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INVITED REVIEW
Modern Techniques for the Isolation of Extracellular Vesicles and Viruses Ryan P. McNamara 1
&
Dirk P. Dittmer 1
Received: 5 March 2019 / Accepted: 15 August 2019 # Springer Science+Business Media, LLC, part of Springer Nature 2019
Abstract Extracellular signaling is pivotal to maintain organismal homeostasis. A quickly emerging field of interest within extracellular signaling is the study of extracellular vesicles (EV), which act as messaging vehicles for nucleic acids, proteins, metabolites, lipids, etc. from donor cells to recipient cells. This transfer of biologically active material within a vesicular body is similar to the infection of a cell through a virus particle, which transfers genetic material from one cell to another to preserve an infection state, and viruses are known to modulate EV. Although considerable heterogeneity exists within EVand viruses, this review focuses on those that are small (< 200 nm in diameter) and of relatively low density (< 1.3 g/mL). A multitude of isolation methods for EV and virus particles exist. In this review, we present an update on methods for their isolation, purification, and phenotypic characterization. We hope that the information we provide will be of use to basic science and clinical investigators, as well as biotechnologists in this emerging field. Keywords Extracellular vesicles . Viruses . Exosomes . Microvesicles
Introduction Extracellular vesicles (EV) are secreted from every cell type studied. Several subtypes of EV exist and are classified based on their sizes, cellular origin and surface markers. Apoptotic bodies (800–5000 nm in diameter) and microvesicles (100– 800 nm in diameter) bud off at the plasma membrane and are thus enriched for proteins sitting on the cell surface. Exosomes are a class of EV of a small diameter (40–150 nm in diameter) that originate from the inward budding of endosomes into the multivesicular body and contain endosomal trafficking markers such as tumor susceptibility gene 101 (Tsg101), apoptosislinked gene-2 interacting protein X (Alix), tetraspanins, and flotillins (reviewed in (Crescitelli et al. 2013; Raab-Traub and Dittmer 2017)). EV have received a substantial amount of attention in recent years. They act as an extracellular mailing system, transferring information from one cell to the next, and
* Dirk P. Dittmer [email protected] 1
Lineberger Comprehensive Cancer Center, Department of Microbiology and Immunology, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA
can contain nucleic acids such as DNA, micro RNAs (miRNA) and non-coding RNAs (ncRNA), mRNA, proteins and enzymes such as histones and esterases, metabolites, and lipids (Chugh et al. 2013; Hurwitz et al. 2016; Meckes et al. 2010, 2013; Wang et al. 2015; Willms et al. 2016; Bukong et al. 2014; Longatti et al. 2015). EV have been proposed to play a critical role in cell differentiation, angiogenesis, metabolic reprogramming, tumor progression, immune modulation, and response to pathogen challenge (Willms et al. 2018;
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