Isolation of Extracellular Vesicles in Saliva Using Density Gradient Ultracentrifugation

Methods Mol Biol. 2017;1660:343-350. doi: 10.1007/978-1-4939-7253-1_27.

Abstract

This chapter describes a method for isolating human salivary extracellular vesicles (EVs) using density gradient ultracentrifugation. Standard protocols established for isolation of EVs from blood or a conditioned medium of cultured cells do not work for whole saliva, due to its viscosity. Therefore, procedures including a pretreatment step and utilizing iodixanol as a gradient material enable EVs to be concentrated to a 1.1 g/ml density. This protocol is compatible with both swing and angle rotors. By employing an angle rotor, which enables high g-force, the centrifugation time was reduced to 4 h from the 17 h required when using a swing rotor.

Keywords: Angle rotor; EVs; Exosome; Extracellular vesicles; Iodixanol; Whole saliva.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Centrifugation, Density Gradient*
  • Extracellular Vesicles / metabolism*
  • Humans
  • Saliva / metabolism*
  • Ultracentrifugation*
  • Workflow