用放射性铬释放法和 DELFIA-EuTDA 细胞毒性检测法分离和检测 NK 细胞的细胞毒性。

NK Cell Isolation and Cytotoxicity by Radioactive Chromium Release Assay and DELFIA-EuTDA Cytotoxicity Assay.

机构信息

Department of Microbiology, Immunology and Genetics, University of North Texas Health Science Center, Fort Worth, TX, USA.

出版信息

Methods Mol Biol. 2022;2463:221-233. doi: 10.1007/978-1-0716-2160-8_16.

DOI:10.1007/978-1-0716-2160-8_16

PMID:35344178

Abstract

Cytotoxicity assays are important in vitro tools to measure the lysis of desired target cells via an effector immune cell of choice. Specific lysis of the target cells can be determined by labeling the target cells with a radioactive isotope or fluorescent molecule, co-incubating it with an effector cell, then measuring the release of the labeled molecule in the supernatant. Here, we describe and compare different cell cytotoxicity assays using a chromium-51 (Cr) release and DELFIA EuTDA fluorescent assay using K562 as the target cells and peripheral blood mononuclear cell (PBMC) derived natural killer (NK) cells as the effector cells.

摘要

细胞毒性测定是一种重要的体外工具，用于通过选择的效应免疫细胞来测量所需靶细胞的裂解。通过用放射性同位素或荧光分子标记靶细胞，将其与效应细胞共孵育，然后测量上清液中标记分子的释放，即可确定靶细胞的特异性裂解。在这里，我们使用铬-51（Cr）释放和 DELFIA EuTDA 荧光测定法描述和比较了不同的细胞毒性测定法，使用 K562 作为靶细胞和外周血单核细胞（PBMC）衍生的自然杀伤（NK）细胞作为效应细胞。