In vitro protocol for the optimal induction of inflammation in human monocyte cell lines

The study of inflammation is of key interest to biomedical research; however, techniques to induce inflammation in vitro are difficult to implement. Here, we present a protocol that optimizes the induction and measurement of NF-kB-mediated inflammation in vitro using a human macrophage cell line. We describe steps for growing, differentiating, and inducing inflammation in THP-1 cells. We detail the process of staining and grid-based confocal imaging. We also discuss approaches to test anti-inflammatory drug efficacy in inhibiting inflammatory milieu. For complete details on the use and execution of this protocol, please refer to Koganti et al. (2022).¹.