Advances in Technologies for Purification and Enrichment of Extracellular Vesicles

Abstract

Extracellular vesicles (EVs) are lipid bilayer-bound vesicles secreted by cells. Subtypes of EVs such as microvesicles and exosomes are further categorized mainly by their different biogenesis mechanisms. EVs have been revealed to play an important role in disease diagnosis and intercellular communication. Despite the wide interest in EVs, the technologies for the purification and enrichment of EVs are still in their infancy. The isolation of EVs, especially exosomes, is inherently challenging due to their small size and heterogeneity. In this review, we mainly introduce the advances of techniques in isolating microvesicles and exosomes according to their approaches. Also, we discuss the limitations of currently reported technologies in terms of their specificity and efficiency, and provide our thoughts about future developments of EV purification and enrichment technology.

Human Oral Stem Cells, Biomaterials and Extracellular Vesicles: A Promising Tool in Bone Tissue Repair

“…Released membrane vesicles from eukaryotic cells, as exosomes, microparticles, microvesicles and apoptotic bodies, can be retained as a dynamic extracellular vesicular compartment, strategic for their paracrine or autocrine biological effects in tissue metabolism [43]. In particular, EVs and their soluble secretory products, also called ‘secretome’, contain abundant proteins, nucleic acids, lipids and a pool of soluble cytokines; they seem to act as biomarkers in many cell functions [44]. Due to their size (50–1000 nm), EVs can circulate in the human body and in particular are present in most biological fluids.…”

“…Specific protein markers for EVs have been planned so that isolation and verification could be easier. In fact, many markers in EVs have been evidenced, such as: Endosomal sorting complexes required for transport (ESCRT), system components (such as TSG101 and Alix) [45], tetraspanin proteins (such as CD9 and CD63) and some Rab small GTPase participating in the biogenesis of EVs (such as Rab27a, Rab27b and Rab35) [44]. Recently, EVs from GMSCs have been characterized through multiparametric analysis and they have been used in functionalizing the scaffold for bone tissue regeneration [9].…”

Extracellular Vesicles Derived from Human Gingival Mesenchymal Stem Cells: A Transcriptomic Analysis

“…The hGMSCs exert their therapeutic effects also through the release of extracellular vesicles (EVs) [18]. The EVs are small membrane vesicles containing abundant proteins, lipids, a pool of soluble cytokines and nucleic acids such as mRNA and microRNA [19]. EVs represent intercellular communication systems able to interact with target cells by binding to the cell surface receptors, transferring membrane proteins, merging their membrane contents into cell recipient cell plasma membrane [20,21].…”

Extracellular Vesicles in Mycobacterial Infections: Their Potential as Molecule Transfer Vectors

“…Acoustic sorting is another novel method based on the movement of particles in an applied acoustic field, where radiation forces proportional to particle volumes drive the faster migration of larger particles. This results in objects of different size separating from each other in different laminar flows (145). Precipitation with highly hydroxylated, water excluding polymers such as polyethylene glycol, dextrans, and polyvinyls is a method traditionally used to isolate viruses which has been adapted to exosome purification (146) and is the basis of the commercial kits ExoQuick (System Biosciences) and Total Exosomes Isolation Kit (Life Technologies).…”