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

doi:10.1016/j.xpro.2023.102365

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

STAR Protoc. 2023 Sep 15;4(3):102365. doi: 10.1016/j.xpro.2023.102365. Epub 2023 Jul 7.

Authors

Evelyn Lattmann¹, Valérie Lapaire², Mitchell P Levesque³

Affiliations

¹ Department of Dermatology, University of Zurich, University Hospital Zurich, Schlieren, ZH 8952, Switzerland. Electronic address: [email protected].
² Department of Dermatology, University of Zurich, University Hospital Zurich, Schlieren, ZH 8952, Switzerland.
³ Department of Dermatology, University of Zurich, University Hospital Zurich, Schlieren, ZH 8952, Switzerland. Electronic address: [email protected].

Abstract

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.

Keywords: Cancer; Cell Biology; Cell culture; Cell separation/fractionation; Flow Cytometry/Mass Cytometry; Health Sciences; Proteomics.

MeSH terms

Chromatography, Gel
Extracellular Vesicles* / chemistry
Humans
Liquid Biopsy
Melanoma* / diagnosis
Melanoma* / metabolism