Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Journal of Extracellular Vesicles, Год журнала: 2024, Номер 13(2)

Опубликована: Фев. 1, 2024

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of cell origin and change functions phenotypes other cells. These features indicate strong biomarker therapeutic potential have generated broad interest, as evidenced by steady year-on-year increase in numbers scientific publications about EVs. Important advances been made EV metrology understanding applying biology. However, hurdles remain to realising EVs domains ranging from basic biology clinical applications due challenges nomenclature, separation non-vesicular extracellular particles, characterisation functional studies. To address opportunities this rapidly evolving field, International Society for Vesicles (ISEV) updates its 'Minimal Information Studies Vesicles', which was first published 2014 then 2018 MISEV2014 MISEV2018, respectively. The goal current document, MISEV2023, is provide researchers with an updated snapshot available approaches advantages limitations production, multiple sources, including culture, body fluids solid tissues. In addition presenting latest art principles research, document also covers advanced techniques that are currently expanding boundaries field. MISEV2023 includes new sections on release uptake a brief discussion vivo study Compiling feedback ISEV expert task forces more than 1000 researchers, conveys research facilitate robust discoveries move field forward even rapidly.

Язык: Английский

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A comprehensive review on recent advances in exosome isolation and characterization: Toward clinical applications

Translational Oncology, Год журнала: 2024, Номер 50, С. 102121 - 102121

Опубликована: Сен. 14, 2024

Язык: Английский

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The role and applications of extracellular vesicles in osteoporosis

Bone Research, Год журнала: 2024, Номер 12(1)

Опубликована: Янв. 23, 2024

Abstract Osteoporosis is a widely observed condition characterized by the systemic deterioration of bone mass and microarchitecture, which increases patient susceptibility to fragile fractures. The intricate mechanisms governing homeostasis are substantially impacted extracellular vesicles (EVs), play crucial roles in both pathological physiological contexts. EVs derived from various sources exert distinct effects on osteoporosis. Specifically, released osteoblasts, endothelial cells, myocytes, mesenchymal stem cells contribute formation due their unique cargo proteins, miRNAs, cytokines. Conversely, secreted osteoclasts immune promote resorption inhibit formation. Furthermore, use as therapeutic modalities or biomaterials for diagnosing managing osteoporosis promising. Here, we review current understanding impact homeostasis, including classification biogenesis regulatory present an overview latest research progress treating using EVs. Finally, discuss challenges prospects translational

Язык: Английский

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Advancements in extracellular vesicles biomanufacturing: a comprehensive overview of large-scale production and clinical research

Frontiers in Bioengineering and Biotechnology, Год журнала: 2025, Номер 13

Опубликована: Фев. 19, 2025

Extracellular vesicles (EVs) are nano-sized, membranous structures secreted by cells into the extracellular space, have attracted considerable attention in field of biosciences for their role intercellular communication various physiological and pathological processes. Their ubiquitous presence bodily fluids cell-specific characteristics make them promising candidates as biomarkers. Additionally, ability to transport biological therapeutics across different barriers specific target underscores significant translational potential diagnostic therapeutic purposes. Significant progress has been achieved translation EVs research clinical applications, however, challenges persist large-scale production EVs, particularly areas scalable manufacturing, efficient isolation methods, drug loading techniques, advanced characterization technology. This review critically examines complex processes involved biogenesis explores recent developments production. By synthesizing knowledge from these fields, this aims provide a holistic perspective on evolving landscape its underscoring both accomplishments obstacles that lie ahead fully realizing biomedicine.

Язык: Английский

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Improving the Purity of Extracellular Vesicles by Removal of Lipoproteins from Size Exclusion Chromatography- and Ultracentrifugation-Processed Samples Using Glycosaminoglycan-Functionalized Magnetic Beads

ACS Applied Materials & Interfaces, Год журнала: 2024, Номер 16(34), С. 44386 - 44398

Опубликована: Авг. 16, 2024

Extracellular vesicles (EVs) are present in blood at much lower concentrations (5-6 orders of magnitude) compared to lipoprotein particles (LP). Because LP and EV overlap size density, isolating high-purity EVs is a significant challenge. While the current two-step sequential isolation process using size-expression chromatography (SEC) followed by density gradient (DG) achieves high purity, time-consuming ultracentrifugation (UC) step DG hinders workflow efficiency. This paper introduces an optimized magnetic bead reagent, LipoMin, functionalized with glycosaminoglycans (GAGs), as rapid alternative for removal during second-step about 10 minutes. We evaluated LipoMin's efficacy on two sample types: (a) fractions isolated exclusion (SEC + LipoMin) (b) pellet obtained from (UC LipoMin). The remarkably simple, involving min incubation LipoMin separation LP-depleted EV-containing supernatant. Results enzyme-linked immunosorbent assay (ELISA) revealed that removes 98.2% ApoB SEC fractions, comparable ability process. Importantly, yield (CD81 ELISA) remained 93.0% Western blot analysis confirmed key markers, flotillin CD81, were not compromised. Recombinant (rEV), reference standard, was spiked into recovered 89% CD81 protein. For UC ApoA1 decreased 76.5% while retaining 90.7% CD81. Notably, both colorectal cancer (CRC) Alzheimer's disease (AD) samples processed displayed clear expression relevant clinical markers. With (resulting 96% time saving traditional method), reagent offers efficient depletion, paving way streamlined

Язык: Английский

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Lipidomic Analysis of Plasma Extracellular Vesicles Derived from Alzheimer’s Disease Patients

Cells, Год журнала: 2024, Номер 13(8), С. 702 - 702

Опубликована: Апрель 18, 2024

Analysis of blood-based indicators brain health could provide an understanding early disease mechanisms and pinpoint possible intervention strategies. By examining lipid profiles in extracellular vesicles (EVs), secreted particles from all cells, including astrocytes neurons, circulating clinical samples, important insights regarding the brain’s composition can be gained. Herein, a targeted lipidomic analysis was carried out EVs derived plasma samples after removal lipoproteins individuals with Alzheimer’s (AD) healthy controls. Differences were observed for selected species glycerolipids (GLs), glycerophospholipids (GPLs), lysophospholipids (LPLs) sphingolipids (SLs) across three distinct EV subpopulations (all-cell origin, by immunocapture CD9, CD81 CD63; neuronal L1CAM; astrocytic GLAST). The findings new into isolated specific families (MG, DG, Cer, PA, PC, PE, PI, LPI, LPE, LPC), as well differences between AD control individuals. This study emphasizes crucial role lipidomics comprehensive approach identifying biomarkers biological targets related disorders, facilitating diagnosis potentially informing novel interventions.

Язык: Английский

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Multimode chromatography‐based techniques for high purity isolation of extracellular vesicles from human blood plasma

Journal of Extracellular Biology, Год журнала: 2024, Номер 3(3)

Опубликована: Март 1, 2024

Abstract Extracellular vesicles (EVs) play a pivotal role in various biological pathways, such as immune responses and the progression of diseases, including cancer. However, it is challenging to isolate EVs at high purity from blood plasma other biofluids due their low abundance compared more predominant biomolecular species lipoprotein particles free protein complexes. Ultracentrifugation‐based EV isolation, current gold standard technique, cannot overcome this challenge similar biophysical characteristics species. We developed several novel approaches enrich while depleting contaminating molecular using multimode chromatography‐based strategies. On average, we identified 716 ± 68 1054 35 groups isolates 100 µL chromatography‐ ultracentrifugation‐based techniques, respectively. The methods resulted purity, providing significant advantages simplicity, throughput, scalability, applicability for downstream analytical potential clinical applications.

Язык: Английский

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Confirming size-exclusion chromatography as a clinically relevant extracellular vesicles separation method from 1mL plasma through a comprehensive comparison of methods

Deleted Journal, Год журнала: 2024, Номер 1(1)

Опубликована: Июль 1, 2024

Abstract Background Extracellular vesicles (EVs) are amongst the most promising candidates for developing blood-based biomarkers. However, patient sample availability is a key barrier to translational research whilst biobanks store samples of 1.5mL volume or less. To date, there no consensus on suitable method EV separation and current techniques frequently require large volumes biofluids, complicated technology, technical expertise, significant operating costs, which prevents their widespread adoption by less EV-focussed laboratories. Therefore, need an easy reproducible that separates representative EVs from clinically relevant 1mL plasma prior subsequent biomarker identification. Methods In this study, were separated human using four different techniques: size exclusion chromatography (SEC), differential ultracentrifugation, precipitation, immunoaffinity magnetic bead capture. The characterised several orthogonal (protein quantification, nanoparticle tracking analysis, transmission electron microscopy, Western blot, single particle interferometric reflectance imaging sensing, mass spectrometry-based proteomics) comprehensively compare samples. Results We provide examples anticipated results highlighting SEC-processed have greater protein quantification yield, yield expected EVs, sufficient purity, facilitates effective cargo assessment proteomics. Moreover, we confirm overlap with known EV-related proteins within Vesiclepedia database. Additionally, sensing (Leprechaun®), identify SEC has surface tetraspanin distribution population compared unprocessed plasma. Discussion Given requires minimal technology can separate 90 min, comparison reinforces as making it implementation.

Язык: Английский

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Minimal Information for Studies of Extracellular Vesicles (MISEV): Ten-Year Evolution (2014–2023)

Pharmaceutics, Год журнала: 2024, Номер 16(11), С. 1394 - 1394

Опубликована: Окт. 29, 2024

In the tenth year since first edition of MISEV was released in 2014, MISEV2023 has been reported 2024 with aim refining standard and improving rigor, reproducibility, transparency extracellular vesicle (EV) research to clarify requirements for experimental design EVs, emphasize importance reproducible results as well encouraging openness information. The release significantly contributed quality field which creates a more reliable environment. However, despite important role MISEV, there is still need EV researchers continue push widespread implementation guidelines meet evolving nature challenges research. evolution attention it receives have grown exponentially over time, number people involved writing MISEV. Here, this review briefly summarizes three editions aiming recall MISEV2014 MISEV2018 while learning about latest release, MISEV2023, gain deeper understanding content, provide quick note beginners who want learn explore world.

Язык: Английский

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Identification of markers for the isolation of neuron-specific extracellular vesicles

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Апрель 3, 2024

Abstract Extracellular vesicles (EVs) are released by all cells and contain RNA protein from their cell of origin. EVs in biofluids could be used as diagnostic biomarkers to non-invasively report the state inaccessible cells, such neurons brain. As cerebrospinal fluid (CSF) plasma originating many different isolating type-specific measuring cargo help determine specific types. Here, we demonstrate an approach aiming immuno-isolate based on neuron-derived surface markers. We first developed a framework select transmembrane proteins suitable neuron-specific EV markers gene expression proteomics data. Leveraging novel, high-purity isolation method developed, further cataloged present human CSF plasma. Using ultrasensitive immunoassays against several predicted proteins, confirmed one marker, NRXN3 size exclusion chromatography (SEC) density gradient centrifugation (DGC). Finally, efficient immuno-isolation methods applied them isolate + EVs. Our study provides general methodology for cell-type paves way use diagnose neurological disease.

Язык: Английский

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