title: Possibilities and limitations of current technologies for quantification of biological extracellular vesicles and synthetic mimics.
abstract: Nano-sized extracelullar vesicles (EVs) released by various cell types play important roles in a plethora of (patho)physiological processes and are increasingly recognized as biomarkers for disease. In addition, engineered EV and EV-inspired liposomes hold great potential as drug delivery systems. Major technologies developed for high-throughput analysis of individual EV include nanoparticle tracking analysis (NTA), tunable resistive pulse sensing (tRPS) and high-resolution flow cytometry (hFC). Currently, there is a need for comparative studies on the available technologies to improve standardization of vesicle analysis in diagnostic or therapeutic settings. We investigated the possibilities, limitations and comparability of NTA, tRPS and hFC for analysis of tumor cell-derived EVs and synthetic mimics (i.e. differently sized liposomes). NTA and tRPS instrument settings were identified that significantly affected the quantification of these particles. Furthermore, we detailed the differences in absolute quantification of EVs and liposomes using the three technologies. This study increases our understanding of possibilities and pitfalls of NTA, tRPS and hFC, which will benefit standardized and large-scale clinical application of (engineered) EVs and EV-mimics in the future.
authors.affiliation: Department of Neurosurgery, University Medical Center Utrecht, The Netherlands; Brain Center Rudolf Magnus, University Medical Center Utrecht, The Netherlands.
authors.forename: Sybren L N
authors.initials: SL
authors.lastname: Maas
authors.affiliation: Department of Neurosurgery, University Medical Center Utrecht, The Netherlands; Brain Center Rudolf Magnus, University Medical Center Utrecht, The Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: de Vrij
authors.affiliation: Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, The Netherlands.
authors.forename: Els J
authors.initials: EJ
authors.lastname: van der Vlist
authors.affiliation: Department of Neurosurgery, University Medical Center Utrecht, The Netherlands; Brain Center Rudolf Magnus, University Medical Center Utrecht, The Netherlands.
authors.forename: Biaina
authors.initials: B
authors.lastname: Geragousian
authors.affiliation: Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Louis
authors.initials: L
authors.lastname: van Bloois
authors.affiliation: Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Enrico
authors.initials: E
authors.lastname: Mastrobattista
authors.affiliation: Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, The Netherlands.
authors.forename: Raymond M
authors.initials: RM
authors.lastname: Schiffelers
authors.affiliation: Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, The Netherlands.
authors.forename: Marca H M
authors.initials: MH
authors.lastname: Wauben
authors.affiliation: Department of Neurosurgery, University Medical Center Utrecht, The Netherlands; Brain Center Rudolf Magnus, University Medical Center Utrecht, The Netherlands.
authors.forename: Marike L D
authors.initials: ML
authors.lastname: Broekman
authors.affiliation: Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, The Netherlands. Electronic address: e.n.m.nolte@uu.nl.
authors.forename: Esther N M
authors.initials: EN
authors.lastname: Nolte-'t Hoen
mesh_terms: D040761:Biomimetic Materials; D045744:Cell Line, Tumor; D055354:Exosomes; D005434:Flow Cytometry; D006801:Humans; D008081:Liposomes / Q000032:analysis*; D053758:Nanoparticles / Q000032:analysis*
chemical_list: D008081:Liposomes
keywords: Exosomes; Extracellular vesicles; High-resolution flow cytometry; Liposomes; Nanoparticle tracking analysis; Tunable resistive pulse sensing