Riedelberger Michael, Kuchler Karl
Medical University of Vienna, Center for Medical Biochemistry, Max Perutz Labs Vienna, Campus Vienna Biocenter, A-1030 Vienna, Austria.
Bio Protoc. 2020 Mar 20;10(6):e3552. doi: 10.21769/BioProtoc.3552.
Tissue-resident macrophages are pivotal for a tightly-regulated iron metabolism at a cellular and systemic level, since subtle iron alterations increase the susceptibility for microbial infections or drive multiple diseases. However, research on cellular iron homeostasis in macrophages remains challenging due to the limited amount of available methods using radioactive Fe isotopes or strong iron chelators, which might be inapplicable in certain experimental settings. This protocol describes the analysis of the quenchable iron pool (QIP) in macrophages by loading these cells with exogenous iron-complexes. Thereby, the cytoplasmic iron pool can be determined, since the iron uptake ability of macrophages inversely correlates with intracellular iron levels. Thus, this assay enables the accurate analysis of even minor alterations in cytoplasmic iron fluxes and is applicable in almost every laboratory environment. In addition, the protocol can also be adopted for other immune cell types and .
组织驻留巨噬细胞对于细胞和全身水平严格调控的铁代谢至关重要，因为铁的细微变化会增加微生物感染的易感性或引发多种疾病。然而，由于使用放射性铁同位素或强铁螯合剂的可用方法数量有限，巨噬细胞中细胞铁稳态的研究仍然具有挑战性，这些方法在某些实验环境中可能不适用。本方案描述了通过用外源性铁复合物加载巨噬细胞来分析其可淬灭铁池（QIP）的方法。由此，可以确定细胞质铁池，因为巨噬细胞的铁摄取能力与细胞内铁水平呈负相关。因此，该检测方法能够准确分析细胞质铁通量中即使是微小的变化，并且适用于几乎所有实验室环境。此外，该方案也可用于其他免疫细胞类型。
