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Membranes
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Progress in Extracellular Vesicle (EV) Analysis
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Progress in Extracellular Vesicle (EV) Analysis

A special issue of Membranes (ISSN 2077-0375). This special issue belongs to the section "Biological Membrane Composition and Structures".

Deadline for manuscript submissions: closed (10 October 2022) | Viewed by 23776

Special Issue Editor

Dr. Krisztina Vukman

E-Mail Website
Guest Editor
Department of Genetics, Cell- and Immunobiology, Semmelweis University, H-1089 Budapest, Hungary
Interests: immunology; mast cells; extracellular vesicles; inflammation; allergy; degranulation
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

We are pleased to invite you to publish a paper (research article, review, communication, etc.) in a Special Issue on “Progress in Extracellular Vesicle (EV) Analysis”.

EVs represent a heterogeneous group of membrane surrounded structures with various sizes and origins. They play a significant role in a broad range of physiological and pathological processes, such as cell-to-cell signaling, antigen presentation, and immunomodulatory activities. EVs participate in various diseases, and their diagnostic and therapeutic exploitation potential is under intense investigation. However, their size and amount often make them difficult to obtain as relatively pure preparations and to characterize them properly. Accordingly, analysis of EVs’ morphology, composition, biogenesis, and function is one of the topics of interest in EV research.

This Special Issue aims to contribute to advancing the current and frontier developments in the field of EV analysis. In this Special Issue, original research articles and reviews are welcome. Research areas may include (but are not limited to) the following: mechanism of EV production, characteristics of EV membranes and internal molecules, novel applications in EV analysis, etc.

We look forward to receiving your contributions.

Dr. Krisztina Vukman
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Membranes is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • extracellular vesicles
  • analysis
  • separation
  • biogenesis
  • morphology
  • membrane content
  • internal molecules
  • mediators
  • cargo

Published Papers (6 papers)

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Research

Jump to: Review

16 pages, 8650 KiB  
Article
Endoplasmin Is a Hypoxia-Inducible Endoplasmic Reticulum-Derived Cargo of Extracellular Vesicles Released by Cardiac Cell Lines
by Anna Koncz, Lilla Turiák, Krisztina Németh, Dorina Lenzinger, Tünde Bárkai, Péter Lőrincz, Helga Zelenyánszki, Krisztina V. Vukman, Edit I. Buzás and Tamás Visnovitz
Membranes 2023, 13(4), 431; https://doi.org/10.3390/membranes13040431 - 13 Apr 2023
Cited by 3 | Viewed by 1781
Abstract
Cardiomyopathies are leading causes of human mortality. Recent data indicate that the cardiomyocyte-derived extracellular vesicles (EVs) released upon cardiac injury are present in circulation. This paper aimed to analyze EVs released under normal and hypoxic conditions by H9c2 (rat), AC16 (human) and HL1 [...] Read more.
Cardiomyopathies are leading causes of human mortality. Recent data indicate that the cardiomyocyte-derived extracellular vesicles (EVs) released upon cardiac injury are present in circulation. This paper aimed to analyze EVs released under normal and hypoxic conditions by H9c2 (rat), AC16 (human) and HL1 (mouse) cardiac cell lines. Small (sEVs), medium (mEVs) and large EVs (lEVs) were separated from a conditioned medium by a combination of gravity filtration, differential centrifugation and tangential flow filtration. The EVs were characterized by microBCA, SPV lipid assay, nanoparticle tracking analysis, transmission and immunogold electron microscopy, flow cytometry and Western blotting. Proteomic profiles of the EVs were determined. Surprisingly, an endoplasmic reticulum chaperone, endoplasmin (ENPL, grp94 or gp96), was identified in the EV samples, and its association with EVs was validated. The secretion and uptake of ENPL was followed by confocal microscopy using GFP-ENPL fusion protein expressing HL1 cells. We identified ENPL as an internal cargo of cardiomyocyte-derived mEVs and sEVs. Based on our proteomic analysis, its presence in EVs was linked to hypoxia in HL1 and H9c2 cells, and we hypothesize that EV-associated ENPL may have a cardioprotective role by reducing cardiomyocyte ER stress. Full article
(This article belongs to the Special Issue Progress in Extracellular Vesicle (EV) Analysis)
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14 pages, 2897 KiB  
Article
Targeting the Structural Integrity of Extracellular Vesicles via Nano Electrospray Gas-Phase Electrophoretic Mobility Molecular Analysis (nES GEMMA)
by Stephanie Steinberger, Sobha Karuthedom George, Lucia Lauková, René Weiss, Carla Tripisciano, Martina Marchetti-Deschmann, Viktoria Weber, Günter Allmaier and Victor U. Weiss
Membranes 2022, 12(9), 872; https://doi.org/10.3390/membranes12090872 - 09 Sep 2022
Cited by 3 | Viewed by 1850
Abstract
Extracellular vesicles (EVs) are in the scientific spotlight due to their potential application in the medical field, ranging from medical diagnosis to therapy. These applications rely on EV stability during isolation and purification—ideally, these steps should not impact vesicle integrity. In this context, [...] Read more.
Extracellular vesicles (EVs) are in the scientific spotlight due to their potential application in the medical field, ranging from medical diagnosis to therapy. These applications rely on EV stability during isolation and purification—ideally, these steps should not impact vesicle integrity. In this context, we investigated EV stability and particle numbers via nano electrospray gas-phase electrophoretic mobility molecular analysis (nES GEMMA) and nanoparticle tracking analysis (NTA). In nES GEMMA, native, surface-dry analytes are separated in the gas-phase according to the particle size. Besides information on size and particle heterogeneity, particle number concentrations are obtained in accordance with recommendations of the European Commission for nanoparticle characterization (2011/696/EU, 18 October 2011). Likewise, and in contrast to NTA, nES GEMMA enables detection of co-purified proteins. On the other hand, NTA, yielding data on hydrodynamic size distributions, is able to relate particle concentrations, omitting electrolyte exchange (and resulting EV loss), which is prerequisite for nES GEMMA. Focusing on EVs of different origin, we compared vesicles concentrations and stability, especially after electrolyte exchange and size exclusion chromatography (SEC). Co-isolated proteins were detected in most samples, and the vesicle amount varied in dependence on the EV source. We found that depletion of co-purified proteins was achievable via SEC, but was associated with a loss of EVs and—most importantly—with decreased vesicle stability, as detected via a reduced nES GEMMA measurement repeatability. Ultimately, we propose the repeatability of nES GEMMA to yield information on EV stability, and, as a result, we propose that nES GEMMA can yield additional valuable information in EV research. Full article
(This article belongs to the Special Issue Progress in Extracellular Vesicle (EV) Analysis)
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20 pages, 2640 KiB  
Article
Extracellular Particles as Carriers of Cholesterol Not Associated with Lipoproteins
by Sergey Landa, Nicolay Verlov, Natalia Fedorova, Mikhail Filatov, Rimma Pantina, Vladimir Burdakov, Elena Varfolomeeva and Vladimir Emanuel
Membranes 2022, 12(6), 618; https://doi.org/10.3390/membranes12060618 - 14 Jun 2022
Cited by 4 | Viewed by 1884
Abstract
Exosomes and exomeres are the smallest microparticles ranging from 20 to 130 nm in diameter. They are found in almost all biological fluids. Exosomes and exomeres are of considerable interest since they can be involved in intercellular signaling and are biological markers of [...] Read more.
Exosomes and exomeres are the smallest microparticles ranging from 20 to 130 nm in diameter. They are found in almost all biological fluids. Exosomes and exomeres are of considerable interest since they can be involved in intercellular signaling and are biological markers of the state of cells, which can be used for diagnostics. The nomenclature of exosomes remains poorly developed. Most researchers try to classify them based on the mode of formation, physicochemical characteristics, and the presence of tetrasporin markers CD9, CD63, and CD81. The data presented in this work show that although exomeres carry tetrasporin biomarkers, they differ from exosomes strongly in lipid composition, especially in cholesterol content. The production of exomeres by cells is associated with the synthesis of cholesterol in cells and is expressed or suppressed by regulators of the synthesis of mevalonate, an intermediate product of cholesterol metabolism. In addition, the work shows that the concentration of extracellular particles in the body correlates with the concentration of cholesterol in the plasma, but weakly correlates with the concentration of cholesterol in lipoproteins. This suggests that not all plasma cholesterol is associated with lipoproteins, as previously thought. Full article
(This article belongs to the Special Issue Progress in Extracellular Vesicle (EV) Analysis)
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Review

Jump to: Research

22 pages, 19031 KiB  
Review
Exosomes: Small Vesicles with Important Roles in the Development, Metastasis and Treatment of Breast Cancer
by Ling’ao Meng, Kedong Song, Shenglong Li and Yue Kang
Membranes 2022, 12(8), 775; https://doi.org/10.3390/membranes12080775 - 12 Aug 2022
Cited by 6 | Viewed by 4202
Abstract
Breast cancer (BC) has now overtaken lung cancer as the most common cancer, while no biopredictive marker isolated from biological fluids has yet emerged clinically. After traditional chemotherapy, with the huge side effects brought by drugs, patients also suffer from the double affliction [...] Read more.
Breast cancer (BC) has now overtaken lung cancer as the most common cancer, while no biopredictive marker isolated from biological fluids has yet emerged clinically. After traditional chemotherapy, with the huge side effects brought by drugs, patients also suffer from the double affliction of drugs to the body while fighting cancer, and they often quickly develop drug resistance after the drug, leading to a poor prognosis. And the treatment of some breast cancer subtypes, such as triple negative breast cancer (TNBC), is even more difficult. Exosomes (Exos), which are naturally occurring extracellular vesicles (EVs) with nanoscale acellular structures ranging in diameter from 40 to 160 nm, can be isolated from various biological fluids and have been widely studied because they are derived from the cell membrane, have extremely small diameter, and are widely involved in various biological activities of the body. It can be used directly or modified to make derivatives or to make some analogs for the treatment of breast cancer. This review will focus on the involvement of exosomes in breast cancer initiation, progression, invasion as well as metastasis and the therapeutic role of exosomes in breast cancer. Full article
(This article belongs to the Special Issue Progress in Extracellular Vesicle (EV) Analysis)
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28 pages, 1875 KiB  
Review
Engineering of MSC-Derived Exosomes: A Promising Cell-Free Therapy for Osteoarthritis
by Jin Cheng, Yixin Sun, Yong Ma, Yingfang Ao, Xiaoqing Hu and Qingyang Meng
Membranes 2022, 12(8), 739; https://doi.org/10.3390/membranes12080739 - 28 Jul 2022
Cited by 18 | Viewed by 5455
Abstract
Osteoarthritis (OA) is characterized by progressive cartilage degeneration with increasing prevalence and unsatisfactory treatment efficacy. Exosomes derived from mesenchymal stem cells play an important role in alleviating OA by promoting cartilage regeneration, inhibiting synovial inflammation and mediating subchondral bone remodeling without the risk [...] Read more.
Osteoarthritis (OA) is characterized by progressive cartilage degeneration with increasing prevalence and unsatisfactory treatment efficacy. Exosomes derived from mesenchymal stem cells play an important role in alleviating OA by promoting cartilage regeneration, inhibiting synovial inflammation and mediating subchondral bone remodeling without the risk of immune rejection and tumorigenesis. However, low yield, weak activity, inefficient targeting ability and unpredictable side effects of natural exosomes have limited their clinical application. At present, various approaches have been applied in exosome engineering to regulate their production and function, such as pretreatment of parental cells, drug loading, genetic engineering and surface modification. Biomaterials have also been proved to facilitate efficient delivery of exosomes and enhance treatment effectiveness. Here, we summarize the current understanding of the biogenesis, isolation and characterization of natural exosomes, and focus on the large-scale production and preparation of engineered exosomes, as well as their therapeutic potential in OA, thus providing novel insights into exploring advanced MSC-derived exosome-based cell-free therapy for the treatment of OA. Full article
(This article belongs to the Special Issue Progress in Extracellular Vesicle (EV) Analysis)
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27 pages, 1594 KiB  
Review
Current Knowledge on Exosome Biogenesis, Cargo-Sorting Mechanism and Therapeutic Implications
by Shenmin Xie, Qin Zhang and Li Jiang
Membranes 2022, 12(5), 498; https://doi.org/10.3390/membranes12050498 - 06 May 2022
Cited by 74 | Viewed by 7742
Abstract
Extracellular vesicles (EVs) are nanoscale membrane vesicles released by donor cells that can be taken up by recipient cells. The study of EVs has the potential to identify unknown cellular and molecular mechanisms in intercellular communication and disease. Exosomes, with an average diameter [...] Read more.
Extracellular vesicles (EVs) are nanoscale membrane vesicles released by donor cells that can be taken up by recipient cells. The study of EVs has the potential to identify unknown cellular and molecular mechanisms in intercellular communication and disease. Exosomes, with an average diameter of ≈100 nanometers, are a subset of EVs. Different molecular families have been shown to be involved in the formation of exosomes and subsequent secretion of exosomes, which largely leads to the complexity of the form, structure and function of exosomes. In addition, because of their low immunogenicity and ability to transfer a variety of bioactive components to recipient cells, exosomes are regarded as effective drug delivery systems. This review summarizes the known mechanisms of exosomes biogenesis, cargo loading, exosomes release and bioengineering, which is of great importance for further exploration into the clinical applications of EVs. Full article
(This article belongs to the Special Issue Progress in Extracellular Vesicle (EV) Analysis)
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