Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids

STAR Protoc. 2020 Dec 22;2(1):100225. doi: 10.1016/j.xpro.2020.100225. eCollection 2021 Mar 19.


We developed a modified protocol, based on differential ultracentrifugation (dUC), to isolate extracellular vesicles and particles (specifically exomeres) (EVPs) from various human and murine sources, including cell lines, surgically resected tumors and adjacent tissues, and bodily fluids, such as blood, lymphatic fluid, and bile. The diversity of these samples requires robust and highly reproducible protocols and refined isolation technology, such as asymmetric-flow field-flow fractionation (AF4). Our isolation protocol allows for preparation of EVPs for various downstream applications, including proteomic profiling. For complete details on the use and execution of this protocol, please refer to Hoshino et al. (2020).

Keywords: Cell Membrane; Cell culture; Cell isolation; Exomeres; Exosomes; Extracellular vesicles and particles; Isolation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Body Fluids / chemistry*
  • Cell Line
  • Centrifugation, Density Gradient*
  • Extracellular Vesicles / chemistry*
  • Fractionation, Field Flow*
  • Humans
  • Mice
  • Proteomics*