STAR Protocols (Sep 2024)

Protocol for isolating extracellular vesicles from pancreatic cancer cells for liver metastatic niche research

  • Anthony Casabianca,
  • Mélanie Bellina,
  • Crissy Dudgeon,
  • Muhammad Sadeqi Nezhad,
  • Chris Harris,
  • Stephany Fiore,
  • Agnes Bernet,
  • Wade Narrow,
  • Orjola Prela,
  • Lan Wang,
  • Igor Astsaturov,
  • Patrick Mehlen,
  • Darren R. Carpizo

Journal volume & issue
Vol. 5, no. 3
p. 103256

Abstract

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Summary: Extracellular vesicles (EVs) are secreted, cell-derived, membrane-bound compartments implicated in various diseases for their ability to influence distant targets and as carriers of biomarkers. Here, we present a protocol for separating EVs from mammalian pancreatic cancer cells and their characterization using western blot and electron microscopy. We then demonstrate how they are utilized to affect tumor development in a murine model of metastatic pancreatic cancer including a method to quantify hepatic tumor burden in histologic samples.For complete details on the use and execution of this protocol, please refer to Dudgeon et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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