对间叶干细胞衍生的EV制剂进行实验室间基于多聚酶珠的表面蛋白分析,确定间叶干细胞-EV表面标记特征。

IF 15.5 1区 医学 Q1 CELL BIOLOGY Journal of Extracellular Vesicles Pub Date : 2024-06-13 DOI:10.1002/jev2.12463
Vivian V. T. Nguyen, Joshua A. Welsh, Tobias Tertel, Andre Choo, Simonides I. van de Wakker, Kyra A. Y. Defourny, Bernd Giebel, Pieter Vader, Jayanthi Padmanabhan, Sai Kiang Lim, Esther N. M. Nolte-'t Hoen, Marianne C. Verhaar, R. Beklem Bostancioglu, Antje M. Zickler, Jia Mei Hong, Jennifer C. Jones, Samir EL Andaloussi, Bas W. M. van Balkom, André Görgens
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引用次数: 0

摘要

间充质基质细胞(MSCs)是一种很有前景的再生疗法,主要通过分泌的细胞外囊泡(EVs)发挥其作用。与细胞产品相比,这些EVs体积小、无生命,更易于处理,而且具有优势。因此,间充质干细胞EVs的治疗潜力正受到越来越多的研究。然而,由于间充质干细胞-EV 制造策略的不同,间充质干细胞-EV 产品应被视为高度多样化。此外,用于间充质干细胞-EV表征的EV表征技术多种多样,这使得可靠的实验室间已发表数据比较变得更加复杂。因此,本研究旨在建立一种通用方法,方便不同间充质干细胞-EV 研究人员对间充质干细胞-EV 制剂进行表征,以促进实验室间的比较。为此,我们使用一种新型的基于多重珠蛋白的EV流式细胞术检测板进行了一次全面的实验室间评估。这项评估涉及五个实验室的 11 种不同间充质干细胞-EV 产品,它们的间充质干细胞来源、培养条件和 EV 制备方法各不相同。通过该检测面板(主要涵盖一系列间充质干细胞相关标志物),我们确定了一组细胞表面标志物,这些标志物在所有探索的间充质干细胞-EV制备的EV上始终呈阳性(CD44、CD73和CD105)或阴性(CD11b、CD45和CD197)。层次聚类分析揭示了与特定制备过程和实验室条件相关的不同表面标记特征。我们建议将 CD73、CD105 和 CD44 作为可靠的阳性标记物,用于最小化鉴定间充质干细胞衍生的 EV;将 CD11b、CD14、CD19、CD45 和 CD79 作为可靠的阴性标记物。此外,我们还强调了培养基成分(尤其是人血小板裂解液)对EV表面标志物特征的影响,突出了培养条件对EV产物的影响。这种可标准化的间充质干细胞-EV 表面标记谱分析方法为临床前和临床研究中制备的 EV 产品的常规表征提供了工具,加强了间充质干细胞-EV 制剂的质量控制,并有望为提高新兴治疗性间充质干细胞-EV 领域的一致性和可重复性铺平道路。
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Inter-laboratory multiplex bead-based surface protein profiling of MSC-derived EV preparations identifies MSC-EV surface marker signatures

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.

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来源期刊
Journal of Extracellular Vesicles
Journal of Extracellular Vesicles Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
27.30
自引率
4.40%
发文量
115
审稿时长
12 weeks
期刊介绍: The Journal of Extracellular Vesicles is an open access research publication that focuses on extracellular vesicles, including microvesicles, exosomes, ectosomes, and apoptotic bodies. It serves as the official journal of the International Society for Extracellular Vesicles and aims to facilitate the exchange of data, ideas, and information pertaining to the chemistry, biology, and applications of extracellular vesicles. The journal covers various aspects such as the cellular and molecular mechanisms of extracellular vesicles biogenesis, technological advancements in their isolation, quantification, and characterization, the role and function of extracellular vesicles in biology, stem cell-derived extracellular vesicles and their biology, as well as the application of extracellular vesicles for pharmacological, immunological, or genetic therapies. The Journal of Extracellular Vesicles is widely recognized and indexed by numerous services, including Biological Abstracts, BIOSIS Previews, Chemical Abstracts Service (CAS), Current Contents/Life Sciences, Directory of Open Access Journals (DOAJ), Journal Citation Reports/Science Edition, Google Scholar, ProQuest Natural Science Collection, ProQuest SciTech Collection, SciTech Premium Collection, PubMed Central/PubMed, Science Citation Index Expanded, ScienceOpen, and Scopus.
期刊最新文献
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