使用 Seahorse 细胞外通量分析仪评估人类自然杀伤细胞代谢变化。

Assessing Changes in Human Natural Killer Cell Metabolism Using the Seahorse Extracellular Flux Analyzer.

机构信息

Department for Molecular Biology, Center for Molecular Biology Severo Ochoa, Spanish National Research Council-Autonomous University of Madrid (CSIC-UAM), Madrid, Spain.

Lymphoid Malignancies Branch, Center for Cancer Research, National Cancer Institute (NIH), Bethesda, MD, USA.

出版信息

Methods Mol Biol. 2022;2463:165-180. doi: 10.1007/978-1-0716-2160-8_12.

DOI:10.1007/978-1-0716-2160-8_12

PMID:35344174

Abstract

Natural killer (NK) cells are cytotoxic cells that mediate anti-tumor and anti-viral immunity. The response of NK cells to different cytokines and stimuli may involve cell survival, proliferation, and changes in their cytotoxic function. These responses will be supported by changes in cellular metabolism. Therefore, changes in NK metabolic parameters could somehow predict changes in NK cell function and cytotoxicity. In this chapter, we describe a protocol to measure NK cell metabolism in primary human NK cells by using an extracellular flux analyzer. This machine measures pH and oxygen changes in the medium and allows the study of NK cell glycolysis and mitochondrial respiration in real time with a small number of cells.

摘要

自然杀伤 (NK) 细胞是具有细胞毒性的细胞，可介导抗肿瘤和抗病毒免疫。NK 细胞对不同细胞因子和刺激物的反应可能涉及细胞存活、增殖和细胞毒性功能的变化。这些反应将得到细胞代谢变化的支持。因此，NK 代谢参数的变化可能在某种程度上预测 NK 细胞功能和细胞毒性的变化。在本章中，我们描述了一种使用细胞外通量分析仪测量原代人 NK 细胞代谢的方案。该仪器测量培养基中 pH 和氧的变化，并允许使用少量细胞实时研究 NK 细胞的糖酵解和线粒体呼吸。