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High-dimensional functional phenotyping of preclinical human CAR T cells using mass cytometry

Michelozzi, Ilaria M; Sufi, Jahangir; Adejumo, Thomas A; Amrolia, Persis J; Tape, Christopher J; Giustacchini, Alice; (2022) High-dimensional functional phenotyping of preclinical human CAR T cells using mass cytometry. STAR Protocols , 3 (1) , Article 101174. 10.1016/j.xpro.2022.101174. Green open access

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Abstract

Here, we present a comprehensive protocol for the generation and functional characterization of chimeric antigen receptor (CAR) T cells and their products by mass cytometry in a reproducible and scalable manner. We describe the production of CAR T cells from human peripheral blood mononuclear cells. We then detail a three-step staining protocol with metal-labeled antibodies and the subsequent mass cytometry analysis. This protocol allows simultaneous characterization of CAR T cell intracellular signaling, activation, proliferation, cytokine production, and phenotype in a single assay.

Type: Article
Title: High-dimensional functional phenotyping of preclinical human CAR T cells using mass cytometry
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.xpro.2022.101174
Publisher version: https://doi.org/10.1016/j.xpro.2022.101174
Language: English
Additional information: © 2022 The Authors. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Infection, Immunity and Inflammation Dept
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
URI: https://discovery.ucl.ac.uk/id/eprint/10143508
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