Alternative Methods for Characterization of Extracellular Vesicles

Abstract

Extracellular vesicles (ECVs) are nano-sized vesicles released by all cells in vitro as well as in vivo. Their role has been implicated mainly in cell–cell communication, but also in disease biomarkers and more recently in gene delivery. They represent a snapshot of the cell status at the moment of release and carry bioreactive macromolecules such as nucleic acids, proteins, and lipids. A major limitation in this emerging new field is the availability/awareness of techniques to isolate and properly characterize ECVs. The lack of gold standards makes comparing different studies very difficult and may potentially hinder some ECVs-specific evidence. Characterization of ECVs has also recently seen many advances with the use of Nanoparticle Tracking Analysis, flow cytometry, cryo-electron microscopy instruments, and proteomic technologies. In this review, we discuss the latest developments in translational technologies involving characterization methods including the facts in their support and the challenges they face.

Extracellular Vesicles of the Hyperthermophilic Archaeon “Thermococcus onnurineus” NA1 T

“…5% due to a detection limit for particles smaller than 100 nm. This drawback of the qNano method was also pointed out by Momen-Heravi et al (20).…”

“…The qNano system (Izon) is a relatively new technology that allows the detection of EVs passing through a nanopore by way of single-molecule electrophoresis (20). The technology is based on the Coulter principle at the nano scale and operates by detecting transient changes in the ionic current generated by the transport of the target particles through a size-tunable nanopore in a polyurethane membrane.…”

Exosomes and Cardiac Repair After Myocardial Infarction

“…Further characterization of isolated exosomes requires complementary biochemical (immunoblotting and flowcytometry), mass spectrometry, and imaging techniques. 64 …”

Microencapsulation technology by nature: Cell derived extracellular vesicles with therapeutic potential

“…Also, scanning electron microscopy [22], single particle electron microscopy [23], and atomic force microscopy were used successfully to visualize individual EVs [13,22]. Further non-conventional techniques of analysis (suitable for the analysis of EVs and liposomes) include dynamic light scattering analysis (DLS), nanoparticle tracking analysis (NTA) [24,25], and fl uorescence nanoparticle tracking analysis (F-NTA) [26], Raman spectroscopy-based techniques, stimulated emission depletion (STED) microscopy, impedance-based fl ow cytometry, and resistive pulse sensing [27]. Presumably, the increasing demands of this research fi eld will boost the development of further specifi c methodologies and user-friendly laboratory instruments fi tted to the size range of EVs.…”