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通过尺寸排阻色谱法从不同生物流体中分离细胞外囊泡。

Extracellular-Vesicle Isolation from Different Biological Fluids by Size-Exclusion Chromatography.

作者信息

Monguió-Tortajada Marta, Morón-Font Miriam, Gámez-Valero Ana, Carreras-Planella Laura, Borràs Francesc E, Franquesa Marcella

机构信息

REMAR-IVECAT Group, Germans Trias i Pujol Health Science Research Institute, Can Ruti Campus, Badalona, Spain.

Department of Cell Biology, Physiology and Immunology, Autonomous University of Barcelona, Bellaterra, Cerdanyola del Vallès, Spain.

出版信息

Curr Protoc Stem Cell Biol. 2019 Jun;49(1):e82. doi: 10.1002/cpsc.82. Epub 2019 Jan 30.

Abstract

This unit describes how to isolate extracellular vesicles (EVs) from different biological fluids using size-exclusion chromatography (SEC) and how to prepare your starting sample and the EV product for downstream applications. EVs are membrane nanovesicles with specific content that reflects the phenotype and functions of the cell of origin, including protected proteins, lipids, metabolites, and nucleic acids. EVs are thus an excellent resource for noninvasive biomarker discovery in a number of pathological situations and are a promising nanotherapeutic tool to overcome the disadvantages associated with cellular therapy. However, there are still no standardized methods to isolate pure EV preparations, as many approaches do not guarantee proper EV purification, free of contaminating non-EV molecules. Currently, SEC is one of the most promising approaches to purify EVs from any biological fluid, as it avoids co-isolation of contaminants and is user friendly and scalable. © 2019 by John Wiley & Sons, Inc.

摘要

本单元介绍了如何使用尺寸排阻色谱法(SEC)从不同生物流体中分离细胞外囊泡(EVs),以及如何为下游应用制备起始样品和EV产品。EVs是具有特定内容物的膜纳米囊泡,其反映了起源细胞的表型和功能,包括受保护的蛋白质、脂质、代谢物和核酸。因此,在许多病理情况下,EVs是用于无创生物标志物发现的极佳资源,并且是一种有前景的纳米治疗工具,可克服与细胞治疗相关的缺点。然而,目前仍没有标准化的方法来分离纯的EV制剂,因为许多方法不能保证正确纯化EV,使其不含污染性的非EV分子。目前,SEC是从任何生物流体中纯化EVs最有前景的方法之一,因为它避免了污染物的共分离,并且用户友好且可扩展。© 2019约翰威立父子出版公司

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