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

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)...

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Detalles Bibliográficos
Autores principales: Lattmann, Evelyn, Lapaire, Valérie, Levesque, Mitchell P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10339257/
https://www.ncbi.nlm.nih.gov/pubmed/37421613
http://dx.doi.org/10.1016/j.xpro.2023.102365
Descripción
Sumario: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.

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