STAR Protocols (Sep 2023)

Isolation and detection of extracellular vesicles from melanoma cells and liquid biopsies using size-exclusion chromatography and nano-flow cytometry

  • Evelyn Lattmann,
  • Valérie Lapaire,
  • Mitchell P. Levesque

Journal volume & issue
Vol. 4, no. 3
p. 102365

Abstract

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Summary: Characterization of extracellular vesicles (EVs) holds great promise for biomarker discovery and understanding of diseases, including melanoma, the deadliest skin cancer type. Here, we describe a size-exclusion chromatography method to isolate and concentrate EVs from patient material including (1) patient-derived melanoma cell line supernatants and (2) plasma and serum biopsies. Additionally, we provide a protocol to analyze EVs by nano-flow cytometry. EV suspensions obtained with the presented protocol can be used for several downstream analyses including RNA sequencing and proteomics. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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