STAR Protocols (Sep 2024)

Protocol to isolate and characterize pulmonary-specific extracellular vesicles in mice

  • Nayoung Lee,
  • Dohyun Kim,
  • Huijae Cho,
  • Gyeongmin Jeong,
  • Seon-Jin Lee,
  • Heedoo Lee

Journal volume & issue
Vol. 5, no. 3
p. 103183

Abstract

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Summary: Extracellular vesicles (EVs) are membranous nanoparticles classified based on their size and surface markers, which can be specific to various cell origins. Here, we present a protocol for the isolation of pulmonary-specific EVs in mice. We describe steps for differential centrifugation, density gradient centrifugation, and commercially available polyethylene glycol(PEG)-based precipitation, employing pulmonary-specific EV-bound chemicals and antibodies. We then detail procedures for the characterization of these EVs through nanoparticle tracking analysis, flow cytometry, scanning electron microscopy, and transmission electron microscopy.For complete details on the use and execution of this protocol, please refer to Lee et al.1,2,3,4 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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