In vitro assays to evaluate CAR-T cell cytotoxicity.

4区生物学 Q4 Biochemistry, Genetics and Molecular Biology

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-10-24 DOI:10.1016/bs.mcb.2023.05.009

Miao Cao, Robert D Carlson, Ross E Staudt, Adam E Snook

引用次数: 0

Abstract

This chapter introduces four commonly used in vitro chimeric antigen receptor (CAR)-T cell cytotoxicity assays (lactate dehydrogenase release assay, ⁵¹Cr 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.

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评估 CAR-T 细胞细胞毒性的体外试验。

本章介绍了四种常用的体外嵌合抗原受体（CAR）-T细胞细胞毒性检测方法（乳酸脱氢酶释放检测、51Cr释放检测、IncuCyte活细胞杀伤检测和xCELLigence实时分析），并提供了xCELLigence实时分析的详细方案。本章以体外检测为重点，首先解释了T细胞诱导细胞毒性的机制，并讨论了如何利用每种检测方法量化T细胞诱导的细胞毒性。由于xCELLigence实时分析法具有高通量定量和简单明了的工作流程，因此本章提供了一个包含试剂和设备、3天分步操作流程和数据分析说明的方案。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Methods in cell biology 生物-细胞生物学

CiteScore

3.10

自引率

0.00%

发文量

125

审稿时长

3 months

期刊介绍： For over fifty years, Methods in Cell Biology has helped researchers answer the question "What method should I use to study this cell biology problem?" Edited by leaders in the field, each thematic volume provides proven, state-of-art techniques, along with relevant historical background and theory, to aid researchers in efficient design and effective implementation of experimental methodologies. Over its many years of publication, Methods in Cell Biology has built up a deep library of biological methods to study model developmental organisms, organelles and cell systems, as well as comprehensive coverage of microscopy and other analytical approaches.