Protocol to study inflammasome activation in induced pluripotent stem cell-derived macrophages

Macrophages are widely used to study inflammasome activity, but current in vitro macrophage models have various limitations. Here, we describe a protocol to differentiate induced pluripotent stem cell (iPSC)-derived macrophages (iMacs) and to characterize these cells via flow cytometry, phagocytosis assays, and whole-cell proteomics. We then describe how to activate a range of different inflammasomes within these cells and assess the inflammasome response through measurement of pyroptosis, cytokine release, ASC speck formation, and protein processing via western blotting. For complete details on the use and execution of this protocol, please refer to McKee et al.¹.