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

Journal of Extracellular Vesicles, Journal Year: 2024, Volume and Issue: 13(2)

Published: Feb. 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.

Language: Английский

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Extracellular Vesicle Preparation and Analysis: A State‐of‐the‐Art Review

Advanced Science, Journal Year: 2024, Volume and Issue: 11(30)

Published: June 14, 2024

Abstract In recent decades, research on Extracellular Vesicles (EVs) has gained prominence in the life sciences due to their critical roles both health and disease states, offering promising applications diagnosis, drug delivery, therapy. However, inherent heterogeneity complex origins pose significant challenges preparation, analysis, subsequent clinical application. This review is structured provide an overview of biogenesis, composition, various sources EVs, thereby laying groundwork for a detailed discussion contemporary techniques preparation analysis. Particular focus given state‐of‐the‐art technologies that employ microfluidic non‐microfluidic platforms EV processing. Furthermore, this discourse extends into innovative approaches incorporate artificial intelligence cutting‐edge electrochemical sensors, with particular emphasis single proposes current outlines prospective avenues future research. The objective motivate researchers innovate expand methods analysis fully unlocking biomedical potential.

Language: Английский

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A systematic review and meta‐analysis of clinical trials assessing safety and efficacy of human extracellular vesicle‐based therapy

Journal of Extracellular Vesicles, Journal Year: 2024, Volume and Issue: 13(7)

Published: July 1, 2024

Abstract Nowadays, it has become clear that extracellular vesicles (EVs) are not a cellular waste disposal vesicle but an essential part of intercellular communication system. Besides the use EVs in biomarker studies and diagnostics, potential EV‐therapeutics been seen by many. They provide unique properties for disease therapy, including strong immune‐modulatory actions, possibility engineering, low immunogenicity, capability crossing biological barriers. Proof‐of‐concept various pathologies achieved preclinical studies. However, clinical trials with have only emerging slowly. Here, we aim to comprehensive overview current state‐of‐the‐art concerning using human therapy. By approaching knowledge systematic manner, were able include 21 reports meta‐analysis safety evaluation efficacy outcomes. Overall, shown EV‐based therapy is safe incidence serious adverse events (SAE; 0.7% (95%‐CI: 0.1–5.2%), (AE; 4.4% 0.7–22.2%). Subgroup analysis showed no significant difference SAE when comparing autologous versus allogeneic administration, as well engineered non‐engineered EV products. A significantly higher number AE was administration. relevance remains questionable. Evaluation outcomes immunostimulatory, immunosuppressive or regenerative EV‐therapies indicated improvement majority treated patients. Despite these promising results, data need be approached caution due high heterogeneity manufacturing methods, study design, reporting (S)AE. conclude presents opportunity More efforts needed standardization harmonization isolation characterization (S)AE allow inter‐study comparison.

Language: Английский

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Mesenchymal stromal cells-derived extracellular vesicles in cartilage regeneration: potential and limitations

Stem Cell Research & Therapy, Journal Year: 2025, Volume and Issue: 16(1)

Published: Jan. 23, 2025

Articular cartilage injuries can lead to pain, stiffness, and reduced mobility, may eventually progress osteoarthritis (OA). Despite substantial research efforts, effective therapies capable of regenerating are still lacking. Mesenchymal stromal cells (MSCs) known for their differentiation immunomodulatory capabilities, yet challenges such as limited survival post-injection inconsistent therapeutic outcomes hinder clinical application. Recent evidence suggests that the beneficial effects MSCs largely mediated by secreted small extracellular vesicles (sEVs), which have been shown promote tissue repair reduce inflammation. MSC-derived sEVs promise in mitigating degradation chondrocyte apoptosis, positioning them a promising alternative MSC-based OA treatment. This review explores potential limitations regeneration. systematic was conducted following PRISMA guidelines, with comprehensive search Web Science Scopus databases studies published between 2019 2024. A total 223 records were identified, 132 articles assessed eligibility based on general selection criteria. After full-text screening, 60 initially selected, comprising 58 vitro 40 vivo studies. Following further exclusion specific criteria, 33 28 from 47 scientific papers included final qualitative synthesis. Most indicate enhance proliferation, improve matrix composition, matrix-degrading enzymes inflammation, thereby delaying progression. growing body supports use tool preventing progression, most reporting structure function. However, remain optimizing standardizing isolation, dosage, delivery methods Further is necessary elucidate mechanisms underlying sEVs-mediated regeneration facilitate translation into therapies.

Language: Английский

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Inter‐laboratory multiplex bead‐based surface protein profiling of MSC‐derived EV preparations identifies MSC‐EV surface marker signatures

Journal of Extracellular Vesicles, Journal Year: 2024, Volume and Issue: 13(6)

Published: June 1, 2024

Abstract Mesenchymal stromal cells (MSCs) are promising regenerative therapeutics that primarily exert their effects through secreted extracellular vesicles (EVs). These EVs – being small and non‐living easier to handle possess advantages over cellular products. Consequently, the therapeutic potential of MSC‐EVs is increasingly investigated. However, due variations in MSC‐EV manufacturing strategies, products should be considered as highly diverse. Moreover, diverse array EV characterisation technologies used for further complicates reliable interlaboratory comparisons published data. this study aimed establish a common method can easily by various researchers characterise preparations facilitate comparisons. To end, we conducted comprehensive inter‐laboratory assessment using novel multiplex bead‐based flow cytometry assay panel. This involved 11 different from five laboratories with varying MSC sources, culture conditions, preparation methods. Through panel covering range mostly MSC‐related markers, identified set cell surface markers consistently positive (CD44, CD73 CD105) or negative (CD11b, CD45 CD197) on all explored preparations. Hierarchical clustering analysis revealed distinct marker profiles associated specific processes laboratory conditions. We propose CD73, CD105 CD44 robust minimally identifying MSC‐derived CD11b, CD14, CD19, CD79 markers. Additionally, highlight influence medium components, particularly human platelet lysate, profiles, underscoring conditions resulting standardisable approach profiling offers tool routine manufactured pre‐clinical clinical research, enhances quality control preparations, hopefully paves way higher consistency reproducibility emerging field.

Language: Английский

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Proteomic profiling of mesenchymal stem cell‐derived extracellular vesicles: Impact of isolation methods on protein cargo

Journal of Extracellular Biology, Journal Year: 2024, Volume and Issue: 3(6)

Published: June 1, 2024

Abstract Extracellular vesicles (EVs) are nanosized with a lipid bilayer that secreted by cells and play critical role in cell‐to‐cell communication. Despite the promising reports regarding their diagnostic therapeutic potential, utilization of EVs clinical setting is limited due to insufficient information about cargo lack standardization isolation analysis methods. Considering protein cargos as key contributors potency, we conducted tandem mass tag (TMT) quantitative proteomics three subpopulations mesenchymal stem cell (MSC)‐derived obtained through different techniques: ultracentrifugation (UC), high‐speed centrifugation (HS), on sucrose cushion (SU). Subsequently, checked EV marker expression, size distribution, morphological characterization, followed bioinformatic analysis. The proteome results revealed these exhibit distinct molecular functional characteristics. choice method impacts isolated isolating EVs. Specifically, (HS) exhibited higher abundance ribosomal mitochondrial proteins. Functional apoptosis assays comparing mitochondria methods HS‐EVs, but not other EVs, induced early cancer cells. On hand, using (SU) (UC) demonstrated proteins primarily involved immune response, cell‐cell interactions extracellular matrix interactions. Our analyses unveil notable disparities associated biological functions among subpopulations, underscoring importance meticulously selecting resultant based intended application.

Language: Английский

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Quantification of urinary podocyte‐derived migrasomes for the diagnosis of kidney disease

Journal of Extracellular Vesicles, Journal Year: 2024, Volume and Issue: 13(6)

Published: June 1, 2024

Abstract Migrasomes represent a recently uncovered category of extracellular microvesicles, spanning diameter range 500 to 3000 nm. They are emitted by migrating cells and harbour diverse array RNAs proteins. can be readily identified in bodily fluids like serum urine, rendering them valuable non‐invasive source for disease diagnosis through liquid biopsy. In this investigation, we introduce streamlined effective approach the capture quantitative assessment migrasomes, employing wheat germ agglutinin (WGA)‐coated magnetic beads flow cytometry (referred as WBFC). Subsequently, examined levels migrasomes urine kidney (KD) patients with podocyte injury healthy volunteers using WBFC. The outcomes unveiled substantial increase urinary podocyte‐derived migrasome concentrations among individuals KD compared counterparts. Notably, were found express an abundant quantity phospholipase A2 receptor (PLA2R) presence PLA2R proteins these holds promise serving natural antigen quantification autoantibodies against afflicted membranous nephropathy. Consequently, our study not only pioneers novel technique isolation but also underscores potential promising biomarker early injury.

Language: Английский

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Process development for the production of mesenchymal stromal cell-derived extracellular vesicles in conventional 2D systems

Cytotherapy, Journal Year: 2024, Volume and Issue: 26(9), P. 999 - 1012

Published: April 27, 2024

BackgroundIn recent years, the importance of extracellular vesicles (EVs) derived from mesenchymal stromal cells (MSCs) has increased significantly. For their widespread use, a standardized EV manufacturing is needed which often includes conventional, static 2D systems. these system critical process parameters need to be determined.MethodsWe studied impact on MSC proliferation, MSC-derived particle production including EVs, EV- and MSC-specific marker expression, functionality in HaCaT cell migration assay.ResultsWe found that culture growth surface media affected MSCs secretory behavior. Interestingly, materials promoted proliferation did not necessarily result most functional particles. In addition, we seeded at 4 × 103 cm−2 produced particles with improved properties compared higher seeding densities. highly proliferative state produce particles, although were significantly more effective promoting migration. The same correlation was when investigating cultivation temperature. A physiological temperature 37°C optimal for yield, it resulted We observed proliferation-associated potential correlations between glucose consumption, enabling estimation final yields.ConclusionsOur findings suggest parameters, must defined prior each individual do require complex expensive equipment, can increase EVs. Integrating into development paves way robust efficient early clinical phases.Graphical abstract

Language: Английский

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Increasing the biomolecular relevance of cell culture practice

Journal of Biomedical Science, Journal Year: 2025, Volume and Issue: 32(1)

Published: Jan. 3, 2025

Abstract The biomolecular relevance of medium supplements is a key challenge affecting cell culture practice. composition commonly used differs from that physiological environment, the validity conclusions drawn in vitro studies. This article discusses advantages and disadvantages common supplements, including context-dependent considerations for supplement selection to improve relevance, especially nanomedicine extracellular vesicle research.

Language: Английский

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Comparative effects of various extracellular vesicle subpopulations derived from clonal mesenchymal stromal cells on cultured fibroblasts in wound healing-related process

The International Journal of Biochemistry & Cell Biology, Journal Year: 2025, Volume and Issue: unknown, P. 106737 - 106737

Published: Jan. 1, 2025

Language: Английский

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