scholarly journals ISOLATION METHODS OF LARGE AND SMALL EXTRACELLULAR VESICLES DERIVED FROM CARDIOVASCULAR PROGENITORS: A COMPARATIVE STUDY

2021 ◽  
Author(s):  
Laura Saludas ◽  
Elisa Garbayo ◽  
Adrián Ruiz-Villalba ◽  
Silvia Hernández ◽  
Pieter Vader ◽  
...  
Keyword(s):  
Comparative Study ◽  
Extracellular Vesicles ◽  
Isolation Methods

scholarly journals Enhanced recovery of CD9-positive extracellular vesicles from human specimens by chelating reagent

2020 ◽  
Author(s):  
Ayako Kurimoto ◽  
Yuki Kawasaki ◽  
Toshiki Ueda ◽  
Tatsutoshi Inuzuka
Keyword(s):  
Extracellular Vesicles ◽  
Western Blotting ◽  
Biological Samples ◽  
Liquid Biopsy ◽  
Enhanced Recovery ◽  
Recovery Efficiency ◽  
Early Diagnostics ◽  
Isolation Methods ◽  
Chelating Reagent

AbstractExtracellular vesicles (EVs) have gained attention as potential targets of early diagnostics and prognosis in the field of liquid biopsy. Despite clinical potentials, the best method to isolate EVs from specimens remains controversial due to low purity, low specificity, and lack of reproducibility with current isolation methods. Here we show that a chelating reagent enhances the recovery efficiency of EVs from crude biological samples by immunoprecipitation using an anti-CD9 antibody. Proteomic and western blotting analyses show that the EVs isolated using the chelating reagent contain a wider variety of proteins than those isolated with PBS.

scholarly journals Emerging role of extracellular vesicles in the regulation of skeletal muscle adaptation

2019 ◽  
Vol 127 (2) ◽  
pp. 645-653 ◽  
Author(s):  
Ivan J. Vechetti
Keyword(s):  
Skeletal Muscle ◽  
Extracellular Vesicles ◽  
Human Body ◽  
Intercellular Communication ◽  
Skeletal Muscle Mass ◽  
Genetic Material ◽  
Muscle Adaptation ◽  
Pathological Conditions ◽  
Isolation Methods

Extracellular vesicles (EVs) were initially characterized as “garbage bags” with the purpose of removing unwanted material from cells. It is now becoming clear that EVs mediate intercellular communication between distant cells through a transfer of genetic material, a process important to the systemic adaptation in physiological and pathological conditions. Although speculative, it has been suggested that the majority of EVs that make it into the bloodstream would be coming from skeletal muscle, since it is one of the largest organs in the human body. Although it is well established that skeletal muscle secretes peptides (currently known as myokines) into the bloodstream, the notion that skeletal muscle releases EVs is in its infancy. Besides intercellular communication and systemic adaptation, EV release could represent the mechanism by which muscle adapts to certain stimuli. This review summarizes the current understanding of EV biology and biogenesis and current isolation methods and briefly discusses the possible role EVs have in regulating skeletal muscle mass.

scholarly journals New multiscale characterisation methodology for effective determination of isolation-structure-function relationship of extracellular vesicles

2021 ◽  
Author(s):  
Thanh Huyen Phan ◽  
Shiva Kamini Divakarla ◽  
Jia Hao Yeo ◽  
Qingyu Lei ◽  
Priyanka Tharkar ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Biological Effects ◽  
Dry Mass ◽  
Experimental Methodology ◽  
Wide Range ◽  
Isolation Methods ◽  
Function Relationship ◽  
Relationship Of

AbstractExtracellular vesicles (EVs) have been lauded as next generation medicines, but very few EV-based therapeutics have progressed to clinical use. Limited clinical translation is largely due to technical barriers that hamper our ability to mass-produce EVs, i.e. to isolate, purify and characterise them effectively. Technical limitations in comprehensive characterisation of EVs leads to unpredicted biological effects of EVs. Here, using a range of optical and non-optical techniques, we showed that the differences in molecular composition of EVs isolated using two isolation methods correlated with the differences in their biological function. Our results demonstrated that the isolation method determines the composition of isolated EVs at single and sub-population levels. Besides the composition, we measured for the first time the dry mass and predicted sedimentation of EVs. These parameters were shown to correlate well with the biological and functional effects of EVs on single cell and cell cultures. We anticipate that our new multiscale characterisation approach, which goes beyond traditional experimental methodology, will support fundamental understanding of EVs as well as elucidate the functional effects of EVs in in vitro and in vivo studies. Our findings and methodology will be pivotal for developing optimal isolation methods and establishing EVs as mainstream therapeutics and diagnostics. This innovative approach is applicable to a wide range of sectors including biopharma and biotechnology as well as to regulatory agencies.

scholarly journals Methodological guidelines for isolation and purification of plant extracellular vesicles

2021 ◽  
Author(s):  
Yifan Huang ◽  
Shumei Wang ◽  
Qiang Cai ◽  
Hailing Jin
Keyword(s):  
Arabidopsis Thaliana ◽  
Extracellular Vesicles ◽  
Isolation And Purification ◽  
Study Guides ◽  
Pathogen Virulence ◽  
Isolation Methods ◽  
The Cross ◽  
Methodological Guidelines

ABSTRACTPlant extracellular vesicles (EVs) have become the focus of rising interest due to their important roles in the cross-kingdom trafficking of molecules from hosts to interacting microbes to modulate pathogen virulence. However, the isolation of pure intact EVs from plants still represents a considerable challenge. Currently, plant EVs have been isolated from apoplastic washing fluid (AWF) using a variety of methods. Here, we compare two published methods used for isolating plant EVs, and provide a detailed recommended method for AWF collection from Arabidopsis thaliana, followed by EV isolation via differential ultracentrifugation. To further separate and purify specific subclasses of EV from heterogeneous vesicles, sucrose or iodixanol density-based separation and immunoaffinity capture are then utilized. We found that immunoaffinity capture provides a significant advantage for specific EV isolation when suitable specific EV biomarkers and their corresponding antibodies are available. Overall, this study guides the selection and optimization of EV isolation methods for desired downstream applications.

scholarly journals Extracellular Vesicles as an Efficient and Versatile System for Drug Delivery

Cells ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 2191 ◽  
Author(s):  
Xuan T. T. Dang ◽  
Jayasinghe Migara Kavishka ◽  
Daniel Xin Zhang ◽  
Marco Pirisinu ◽  
Minh T. N. Le
Keyword(s):  
Extracellular Vesicles ◽  
Drug Carriers ◽  
Clinical Applications ◽  
Clinical Translation ◽  
Future Perspective ◽  
Target Specificity ◽  
Intrinsic Properties ◽  
Isolation Methods ◽  
Promising Solution ◽  
Major Factors

Despite the recent advances in drug development, the majority of novel therapeutics have not been successfully translated into clinical applications. One of the major factors hindering their clinical translation is the lack of a safe, non-immunogenic delivery system with high target specificity upon systemic administration. In this respect, extracellular vesicles (EVs), as natural carriers of bioactive cargo, have emerged as a promising solution and can be further modified to improve their therapeutic efficacy. In this review, we provide an overview of the biogenesis pathways, biochemical features, and isolation methods of EVs with an emphasis on their many intrinsic properties that make them desirable as drug carriers. We then describe in detail the current advances in EV therapeutics, focusing on how EVs can be engineered to achieve improved target specificity, better circulation kinetics, and efficient encapsulation of therapeutic payloads. We also identify the challenges and obstacles ahead for clinical translation and provide an outlook on the future perspective of EV-based therapeutics.

scholarly journals An Update on Isolation Methods for Proteomic Studies of Extracellular Vesicles in Biofluids

Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3516 ◽  
Author(s):  
Jing Li ◽  
Xianqing He ◽  
Yuanyuan Deng ◽  
Chenxi Yang
Keyword(s):  
Extracellular Vesicles ◽  
Biomarker Discovery ◽  
Future Research ◽  
The Past ◽  
Analysis Strategies ◽  
Isolation Methods ◽  
Specific Proteins ◽  
Therapeutic Tools ◽  
Almost All ◽  
Clinical Potential

Extracellular vesicles (EVs) are lipid bilayer enclosed particles which present in almost all types of biofluids and contain specific proteins, lipids, and RNA. Increasing evidence has demonstrated the tremendous clinical potential of EVs as diagnostic and therapeutic tools, especially in biofluids, since they can be detected without invasive surgery. With the advanced mass spectrometry (MS), it is possible to decipher the protein content of EVs under different physiological and pathological conditions. Therefore, MS-based EV proteomic studies have grown rapidly in the past decade for biomarker discovery. This review focuses on the studies that isolate EVs from different biofluids and contain MS-based proteomic analysis. Literature published in the past decade (2009.1–2019.7) were selected and summarized with emphasis on isolation methods of EVs and MS analysis strategies, with the aim to give an overview of MS-based EV proteomic studies and provide a reference for future research.

scholarly journals Challenges in the Isolation and Proteomic Analysis of Cancer Exosomes—Implications for Translational Research

Proteomes ◽  
2019 ◽  
Vol 7 (2) ◽  
pp. 22 ◽  
Author(s):  
Jadwiga Jablonska ◽  
Monika Pietrowska ◽  
Sonja Ludwig ◽  
Stephan Lang ◽  
Basant Kumar Thakur
Keyword(s):  
Extracellular Vesicles ◽  
Proteome Analysis ◽  
The Body ◽  
Cell Of Origin ◽  
Proteomics Data ◽  
Therapeutic Approaches ◽  
Protein Targets ◽  
Isolation Methods ◽  
Health And Disease ◽  
Novel Protein

Exosomes belong to the group of extracellular vesicles (EVs) that derive from various cell populations and mediate intercellular communication in health and disease. Like hormones or cytokines, exosomes released by cells can play a potent role in the communication between the cell of origin and distant cells in the body to maintain homeostatic or pathological processes, including tumorigenesis. The nucleic acids, and lipid and protein cargo present in the exosomes are involved in a myriad of carcinogenic processes, including cell proliferation, tumor angiogenesis, immunomodulation, and metastasis formation. The ability of exosomal proteins to mediate direct functions by interaction with other cells qualifies them as tumor-specific biomarkers and targeted therapeutic approaches. However, the heterogeneity of plasma-derived exosomes consistent of (a) exosomes derived from all kinds of body cells, including cancer cells and (b) contamination of exosome preparation with other extracellular vesicles, such as apoptotic bodies, makes it challenging to obtain solid proteomics data for downstream clinical application. In this manuscript, we review these challenges beginning with the choice of different isolation methods, through the evaluation of obtained exosomes and limitations in the process of proteome analysis of cancer-derived exosomes to identify novel protein targets with functional impact in the context of translational oncology.

scholarly journals Valorization of Deproteinated Potato Juice Water into β-Glucan Preparation of C. utilis Origin: Comparative Study of Preparations Obtained by Two Isolation Methods

2019 ◽  
Vol 11 (7) ◽  
pp. 3257-3271 ◽  
Author(s):  
Anna Bzducha-Wróbel ◽  
Piotr Koczoń ◽  
Stanisław Błażejak ◽  
Jakub Kozera ◽  
Marek Kieliszek
Keyword(s):  
Comparative Study ◽  
Isolation Methods ◽  
Deproteinated Potato Juice Water

Urinary extracellular vesicles and the kidney: biomarkers and beyond

2014 ◽  
Vol 306 (11) ◽  
pp. F1251-F1259 ◽  
Author(s):  
Mahdi Salih ◽  
Robert Zietse ◽  
Ewout J. Hoorn
Keyword(s):  
Extracellular Vesicles ◽  
Kidney Diseases ◽  
Cell Communication ◽  
Kidney Injury ◽  
Enzyme Linked Immunosorbent Assay ◽  
Polycystic Kidney ◽  
Kidney Physiology ◽  
Isolation Methods ◽  
Biomarker Research ◽  
Potential Use

Extracellular vesicles have been isolated in various body fluids, including urine. The cargo of urinary extracellular vesicles (uEVs) is composed of proteins and nucleic acids reflecting the physiological and possibly pathophysiological state of cells lining the nephron. Because urine is a noninvasive and readily available biofluid, the discovery of uEVs has opened a new field of biomarker research. Their potential use as diagnostic, prognostic, or therapeutic biomarkers for various kidney diseases, including glomerulonephritis, acute kidney injury, tubular disorders, and polycystic kidney disease, is currently being explored. Some challenges, however, remain. These challenges include the need to standardize isolation methods, normalization between samples, and validation of candidate biomarkers. Also, the development of a high-throughput platform to isolate and analyze uEVs, for example, an enzyme-linked immunosorbent assay, is desirable. Here, we review recent studies on uEVs dealing with kidney physiology and pathophysiology. Furthermore, we discuss new and exciting developments regarding vesicles, including their role in cell-to-cell communication and the possibility of using vesicles as a therapy for kidney disorders.

Sign in / Sign up

Export Citation Format

Share Document