A Method for Isolating and Characterizing Mesenchymal Stromal Cell-derived Extracellular Vesicles

Q2 Biochemistry, Genetics and Molecular Biology

Current Protocols in Stem Cell Biology Pub Date : 2018-06-11 DOI:10.1002/cpsc.55

Claudia Lo Sicco, Daniele Reverberi, Luisa Pascucci, Roberta Tasso

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引用次数: 6

Abstract

The unit describes protocols for isolating and characterizing extracellular vesicles (EVs) derived from human adipose tissue-derived mesenchymal stromal cells (MSCs). EVs are a mixed population of membrane-surrounded structures with overlapping composition and size. Advances made in recent years have led to a better understanding of the biological role of EVs. In particular, they can be considered key factors responsible for MSC-paracrine activity, mediating their anti-inflammatory effects towards innate immune cells, such as macrophages.

The topics comprise description of the MSC-conditioned medium containing vesicles preparation, EV isolation, and characterization mainly by specifically set up flow cytometry and electron microscopy approaches, and in vitro methodologies involved in testing the EV anti-inflammatory capacity. The procedures described here can be easily reproduced and can be employed regardless of the type of progenitor cells used to secrete EVs. © 2018 by John Wiley & Sons, Inc.

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间充质间质细胞来源的细胞外囊泡的分离和表征方法

本单元描述了分离和表征来自人脂肪组织来源的间充质基质细胞(MSCs)的细胞外囊泡(ev)的方案。电动汽车是一种混合种群的膜包围结构，具有重叠的组成和大小。近年来取得的进展使人们对电动汽车的生物学作用有了更好的了解。特别是，它们可以被认为是负责msc -旁分泌活性的关键因素，介导它们对先天免疫细胞(如巨噬细胞)的抗炎作用。主题包括描述含有囊泡制备的msc条件培养基，EV分离，主要通过专门设置的流式细胞术和电子显微镜方法进行表征，以及用于测试EV抗炎能力的体外方法。这里描述的过程可以很容易地复制，并且可以用于分泌ev的祖细胞的类型。©2018 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Current Protocols in Stem Cell Biology Biochemistry, Genetics and Molecular Biology-Cell Biology

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期刊介绍： Published in affiliation with the International Society for Stem Cell Research (ISSCR), Current Protocols in Stem Cell Biology (CPSC) covers the most fundamental protocols and methods in the rapidly growing field of stem cell biology. Updated monthly, CPSC will constantly evolve with thelatest developments and breakthroughs in the field. Drawing on the expertise of leading researchers from around the world, Current Protocols in Stem Cell Biology includes methods and insights that will enhance the progress of global research.