A novel extracellular vesicle isolation method based on cellulose nanofiber sheets

Abstract

Extracellular vesicles (EVs) circulate in body fluids, carrying molecular cargo from their parent cells and exerting diverse biological functions. Consequently, they have attracted considerable attention as biomarkers for disease detection and pathophysiological understanding and have emerged as potential therapeutic targets. Although the number of clinical trials involving EVs is increasing, major challenges remain, including methodological transparency and the heterogeneity of EV subpopulations. Various EV isolation methods are commonly employed, and the primary approaches are summarized in the MISEV2023 guidelines. Each method has advantages and disadvantages; however, most conventional approaches require relatively large liquid volumes (e.g., hundreds of microliters or more) to obtain sufficient EV yields for analysis. In recent years, novel technologies have been developed to overcome these limitations by addressing constraints related to sample volume, simplicity, and accuracy. One such innovation is the cellulose nanofiber-EV sheet, which we developed in 2023. This method enables the capture and stable storage of EVs from microvolumes of body fluids (e.g., approximately 10 µL). Two application methods are available: the attaching method, in which the EV sheet is applied to moist tissue surfaces, and the soaking method, in which the sheet is soaked into body fluids. Each method offers distinct advantages. Given their unique properties, EV sheets may contribute to biomarker analysis and facilitate new research directions across diverse fields. Continued advances in EV isolation and analytical platforms will be essential to support the safe clinical implementation of EV-based diagnostics and therapeutics.

Keywords

Extracellular vesicle / EV sheet / cellulose nanofiber / biomarker / ovarian cancer

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