Publication: An integrated toolbox to profile (macrophage) immunometabolism

Publié dans: Cell Reports Methods, 2022, 2 (4), pp.100192. ⟨10.1016/j.crmeth.2022.100192⟩

Auteurs: Sanne G.S. Verberk, Kyra de Goede, Friederike Gorki, Xanthe A.M.H. van Dierendonck, Rafael J Argüello, Jan van den Bossche

Résumé

Macrophages are plastic immune cells that can adopt several activation states. Fundamental to these functional activation states is the regulation of cellular metabolic processes. Especially in mice, the metabolic alterations underlying pro-inflammatory or homeostatic phenotypes have been assessed thoroughly using various techniques. However, researchers new to the field may encounter ambiguity in choosing which combination of techniques is best suited to profile immunometabolism. To guide readers, we provide a toolbox to assess cellular metabolism in a semi-highthroughput 96-well-plate-based format. We applied the approach to activated mouse and human macrophages by metabolically pre-screening cells, followed by measuring extracellular fluxes, mitochondrial mass and membrane potential, glucose and lipid uptake, along with the application of SCENITH. Hereby, we not only validated established activation-induced metabolic rewiring in mouse macrophages, but also uncovered new insights in human macrophage immunometabolism. By thoroughly discussing each techniques, we guide readers with a practical workflow to interrogate immunometabolism.

Lien vers HAL – cnrs-03856610

Lien vers le DOI – 10.1016/j.crmeth.2022.100192