Exosomes from cell culture-conditioned medium: Isolation by ultracentrifugation and characterization

Anurag Purushothaman

Research output: Chapter in Book/Report/Conference proceedingChapter

53 Scopus citations

Abstract

Exosomes are small vesicles of endosomal origin secreted by most cell types. Recent studies have identified exosomes as important mediators of intercellular communication and as important source materials for many clinical applications, including minimal invasive disease diagnosis. Exosomes have been purified from in vitro cell culture supernatants by many different methods; however the most simple and reliable method involves purification by ultracentrifugation. This chapter describes a detailed protocol for isolating exosomes from cell culture-conditioned medium using ultracentrifugation and their characterization based upon size, number, and protein expression by several complementary methods such as transmission electron microscopy, nanoparticle tracking analysis, western blotting, and flow cytometry.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages233-244
Number of pages12
DOIs
StatePublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1952
ISSN (Print)1064-3745

Keywords

  • Cell culture
  • Conditioned medium
  • Exosomes
  • Flow cytometry
  • Sucrose
  • Transmission electron microscopy
  • Ultracentrifugation
  • Western blot

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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