Molecular and Cellular Immunology Section, UCL Great Ormond Street Institute of Child Health, Zayed Centre For Research into Rare Disease in Children, WC1N 1DZ London, UK.
Cell Communication Lab, Department of Oncology, University College London Cancer Institute, WC1E 6DD London, UK.
STAR Protoc. 2022 Feb 9;3(1):101174. doi: 10.1016/j.xpro.2022.101174. eCollection 2022 Mar 18.
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.
在这里，我们提供了一个全面的协议，用于通过质量细胞术以可重复和可扩展的方式生成和功能表征嵌合抗原受体 (CAR) T 细胞及其产物。我们描述了从人外周血单核细胞中生产 CAR T 细胞的过程。然后，我们详细介绍了一个三步染色方案，使用金属标记的抗体，以及随后的质量细胞术分析。该方案允许在单个测定中同时表征 CAR T 细胞的细胞内信号转导、激活、增殖、细胞因子产生和表型。
