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

使用尺寸排阻色谱法和纳米流式细胞术从黑色素瘤细胞和液体活检中分离和检测细胞外囊泡

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作者:Evelyn Lattmann, Valérie Lapaire, Mitchell P Levesque

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.

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