CAMPOS SILVA, CARMEN, Caceres-Martell, Yaiza , Lopez-Cobo, Sheila , Rodriguez, Maria Josefa , Jara, Ricardo , Yanez-Mo, Maria , Vales-Gomez, Mar
Si
Methods Mol. Biol.
Article
Científica
0.368
01/01/2021
000685158600025
Most human cells release extracellular vesicles (EVs) of different sizes and composition, containing biomolecules characteristic from the originating tissue. In consequence, when EVs derive from a cancer cell, they also contain tumor antigens. Therefore, isolating and characterizing tumor-derived EVs has attracted great interest as an invaluable source of biomarkers, both for diagnosis and stratification of cancer. In this chapter, we describe a method for flow cytometry assessment of melanoma-derived EVs which are firstly captured onto antibody-coated beads recognizing either a common EV marker, namely, a tetraspanin, or a tumor antigen like the stress-related molecules MICA or PDL1. Then, after staining with a fluorophore-conjugated antibody directed against a different protein present on the EV surface, the EV-bead complex can be visualized in a conventional flow cytometer. The technique allows detection of proteins present on EVs isolated from tissue culture supernatants of melanoma cell lines and, more importantly, directly from plasma.
Extracellular vesicles; Exosomes; Cancer; Flow cytometry; Liquid biopsy; Tetraspanin; NKG2D-ligand; MICA; PDL1