Exosomes Extraction and Identification

Methods Mol Biol. 2019:2054:81-91. doi: 10.1007/978-1-4939-9769-5_4.

Abstract

Exosomes are small endosome-derived lipid nanoparticles (50-120 nm in diameter), actively secreted by exocytosis in most living cells. Recently, there is a growing interest of research focused on studying the exosome functions and to understand ways to use them for therapeutic applications in a wide variety of disorders, such as cancer, cardiovascular, neurodegenerative, and musculoskeletal diseases. Recently, a number of techniques have been developed for the isolation of exosomes such as ultracentrifugation, micro-filtration centrifugation, gradient centrifugation, and size-exclusion chromatography. In this chapter, we reveal the protocol and key insights into the isolation, purification, and characterization of exosomes using ultracentrifugation method.

Keywords: Exosome; Labeling; Size distribution; TEM; Ultracentrifugation; Western blot.

MeSH terms

  • Animals
  • Blotting, Western / methods
  • Cell Culture Techniques / methods
  • Cells, Cultured
  • Culture Media
  • Exosomes / ultrastructure*
  • Fluorescent Dyes / chemistry
  • Microscopy, Electron, Transmission
  • Organic Chemicals / chemistry
  • Osteoblasts
  • Primary Cell Culture / methods
  • Staining and Labeling / methods*
  • Ultracentrifugation / methods*

Substances

  • Culture Media
  • Fluorescent Dyes
  • Organic Chemicals
  • PKH67