Department of Pharmacology, Physiology, & Cancer Biology, Thomas Jefferson University, Philadelphia, PA, United States.
Department of Pharmacology, Physiology, & Cancer Biology, Thomas Jefferson University, Philadelphia, PA, United States; Department of Microbiology & Immunology, Thomas Jefferson University, Philadelphia, PA, United States; Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, United States.
Methods Cell Biol. 2024;183:303-315. doi: 10.1016/bs.mcb.2023.05.009. Epub 2023 Oct 24.
This chapter introduces four commonly used in vitro chimeric antigen receptor (CAR)-T cell cytotoxicity assays (lactate dehydrogenase release assay, Cr release assay, IncuCyte live cell killing assay, and xCELLigence real-time analysis) and provides a detailed protocol for xCELLigence real-time analysis. Focusing on in vitro assays, this chapter starts with explaining the mechanisms and discussing the utilization of each assay to quantify T-cell-induced cytotoxicity. Due to the high-throughput quantification and straightforward workflow of xCELLigence real-time analysis, a protocol entailing reagents and equipment, a 3-day step-by-step procedure, and instructions for data analysis are provided.
本章介绍了四种常用的体外嵌合抗原受体（CAR）-T 细胞细胞毒性测定法（乳酸脱氢酶释放测定法、Cr 释放测定法、IncuCyte 活细胞杀伤测定法和 xCELLigence 实时分析），并提供了 xCELLigence 实时分析的详细方案。本章侧重于体外测定法，首先解释了每种测定法的机制和用途，以量化 T 细胞诱导的细胞毒性。由于 xCELLigence 实时分析具有高通量定量和简单的工作流程，因此提供了包含试剂和设备、为期 3 天的分步程序以及数据分析说明的方案。
