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Related Experiment Video

Updated: Jun 18, 2025

Isolation of Extracellular Vesicles from Murine Bronchoalveolar Lavage Fluid Using an Ultrafiltration Centrifugation Technique
09:18

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

Nayoung Lee1, Dohyun Kim1, Huijae Cho1

  • 1Department of Biology and Chemistry, Changwon National University, Changwon 51140, South Korea.

STAR Protocols
|August 2, 2024
PubMed
Summary

This study details a protocol for isolating pulmonary-specific extracellular vesicles (EVs) from mice. The method uses advanced techniques for reliable characterization of these important nanoparticles.

Keywords:
Cell BiologyCell isolationFlow CytometryMolecular Biology

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Last Updated: Jun 18, 2025

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Nanomedicine

Background:

  • Extracellular vesicles (EVs) are nanoparticles with cell-specific markers.
  • Isolation of EVs from specific organs like the lungs is crucial for research.
  • Current methods for pulmonary EV isolation require refinement.

Purpose of the Study:

  • To present a detailed protocol for isolating pulmonary-specific extracellular vesicles (EVs) from mice.
  • To enable researchers to obtain pure and well-characterized pulmonary EVs for further study.
  • To establish a reproducible method for pulmonary EV isolation and characterization.

Main Methods:

  • Differential centrifugation and density gradient centrifugation were employed.
  • Polyethylene glycol (PEG)-based precipitation using pulmonary-specific antibodies was utilized.
  • Characterization involved nanoparticle tracking analysis, flow cytometry, and electron microscopy (SEM, TEM).

Main Results:

  • The protocol successfully isolates pulmonary-specific EVs.
  • Characterization methods confirmed the size, surface markers, and morphology of isolated EVs.
  • The described techniques provide a comprehensive approach to pulmonary EV isolation.

Conclusions:

  • This protocol offers a reliable method for obtaining pulmonary-specific EVs.
  • The characterized EVs can be used for various downstream applications in lung research.
  • The study provides a valuable resource for researchers in the field of extracellular vesicles and respiratory diseases.