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

作者信息

Nguyen Vivian V T, Welsh Joshua A, Tertel Tobias, Choo Andre, van de Wakker Simonides I, Defourny Kyra A Y, Giebel Bernd, Vader Pieter, Padmanabhan Jayanthi, Lim Sai Kiang, Nolte-'t Hoen Esther N M, Verhaar Marianne C, Bostancioglu R Beklem, Zickler Antje M, Hong Jia Mei, Jones Jennifer C, El Andaloussi Samir, van Balkom Bas W M, Görgens André

机构信息

Department of Nephrology and Hypertension, UMC Utrecht, Utrecht, The Netherlands.

Translational Nanobiology Section, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.

出版信息

J Extracell Vesicles. 2024 Jun;13(6):e12463. doi: 10.1002/jev2.12463.


DOI:10.1002/jev2.12463
PMID:38868945
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11170075/
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 - are easier to handle and possess advantages over cellular products. Consequently, the therapeutic potential of MSC-EVs is increasingly investigated. However, due to variations in MSC-EV manufacturing strategies, MSC-EV products should be considered as highly diverse. Moreover, the diverse array of EV characterisation technologies used for MSC-EV characterisation further complicates reliable interlaboratory comparisons of published data. Consequently, this study aimed to establish a common method that can easily be used by various MSC-EV researchers to characterise MSC-EV preparations to facilitate interlaboratory comparisons. To this end, we conducted a comprehensive inter-laboratory assessment using a novel multiplex bead-based EV flow cytometry assay panel. This assessment involved 11 different MSC-EV products from five laboratories with varying MSC sources, culture conditions, and EV preparation methods. Through this assay panel covering a range of mostly MSC-related markers, we identified a set of cell surface markers consistently positive (CD44, CD73 and CD105) or negative (CD11b, CD45 and CD197) on EVs of all explored MSC-EV preparations. Hierarchical clustering analysis revealed distinct surface marker profiles associated with specific preparation processes and laboratory conditions. We propose CD73, CD105 and CD44 as robust positive markers for minimally identifying MSC-derived EVs and CD11b, CD14, CD19, CD45 and CD79 as reliable negative markers. Additionally, we highlight the influence of culture medium components, particularly human platelet lysate, on EV surface marker profiles, underscoring the influence of culture conditions on resulting EV products. This standardisable approach for MSC-EV surface marker profiling offers a tool for routine characterisation of manufactured EV products in pre-clinical and clinical research, enhances the quality control of MSC-EV preparations, and hopefully paves the way for higher consistency and reproducibility in the emerging therapeutic MSC-EV field.

摘要
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/a7dc4d7eb777/JEV2-13-e12463-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/6148309a8f32/JEV2-13-e12463-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/567701c67bfd/JEV2-13-e12463-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/5a103cedff73/JEV2-13-e12463-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/c331b6d0a718/JEV2-13-e12463-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/a7dc4d7eb777/JEV2-13-e12463-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/6148309a8f32/JEV2-13-e12463-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/567701c67bfd/JEV2-13-e12463-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/5a103cedff73/JEV2-13-e12463-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/c331b6d0a718/JEV2-13-e12463-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0685/11170075/a7dc4d7eb777/JEV2-13-e12463-g004.jpg

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[1]
Inter-laboratory multiplex bead-based surface protein profiling of MSC-derived EV preparations identifies MSC-EV surface marker signatures.

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引用本文的文献

[1]
TNF-α-preconditioning enhances analgesic efficacy of mesenchymal stem cell-derived extracellular vesicle in neuropathic pain miR-101b-3p targeting Nav1.6.

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[2]
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[3]
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[4]
Extracellular Vesicles and Their Role in Skin Inflammatory Diseases: From Pathogenesis to Therapy.

Int J Mol Sci. 2025-4-18

[5]
Force spectroscopy reveals membrane fluctuations and surface adhesion of extracellular nanovesicles impact their elastic behavior.

Proc Natl Acad Sci U S A. 2025-4-22

[6]
Extracellular Vesicles from Different Mesenchymal Stem Cell Types Exhibit Distinctive Surface Protein Profiling and Molecular Characteristics: A Comparative Analysis.

Int J Mol Sci. 2025-4-4

[7]
Multi-omics analysis of uterine fluid extracellular vesicles reveals a resemblance with endometrial tissue across the menstrual cycle: biological and translational insights.

Hum Reprod Open. 2025-2-24

[8]
Mesenchymal Stromal Cell-Derived Extracellular Vesicles in the Management of Atopic Dermatitis: A Scoping Review of Therapeutic Opportunities and Challenges.

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[9]
Recent Advances in Mass Spectrometry-Based Bottom-Up Proteomics.

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[10]
Mesenchymal stem cell exosome therapy: current research status in the treatment of neurodegenerative diseases and the possibility of reversing normal brain aging.

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本文引用的文献

[1]
Transcriptomic and proteomic profiles of fetal versus adult mesenchymal stromal cells and mesenchymal stromal cell-derived extracellular vesicles.

Stem Cell Res Ther. 2024-3-13

[2]
Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches.

J Extracell Vesicles. 2024-2

[3]
Size matters: Functional differences of small extracellular vesicle subpopulations in cardiac repair responses.

J Extracell Vesicles. 2024-1

[4]
The cellular response to extracellular vesicles is dependent on their cell source and dose.

Sci Adv. 2023-9

[5]
Identification of scaffold proteins for improved endogenous engineering of extracellular vesicles.

Nat Commun. 2023-8-7

[6]
Mesenchymal stromal cell exosomes enhance dental pulp cell functions and promote pulp-dentin regeneration.

Biomater Biosyst. 2023-5-26

[7]
Independent human mesenchymal stromal cell-derived extracellular vesicle preparations differentially attenuate symptoms in an advanced murine graft-versus-host disease model.

Cytotherapy. 2023-8

[8]
Cellular expansion of MSCs: Shifting the regenerative potential.

Aging Cell. 2023-1

[9]
A human kidney and liver organoid-based multi-organ-on-a-chip model to study the therapeutic effects and biodistribution of mesenchymal stromal cell-derived extracellular vesicles.

J Extracell Vesicles. 2022-11

[10]
Identification of storage conditions stabilizing extracellular vesicles preparations.

J Extracell Vesicles. 2022-6

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