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

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

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Article numbere55
JournalCurrent Protocols in Stem Cell Biology
Volume46
Issue number1
DOIs
Publication statusPublished - Aug 1 2018
Externally publishedYes

Keywords

  • extracellular vesicles
  • inflammation
  • macrophages
  • mesenchymal stromal cells

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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