Protocol for the generation and assessment of functional macrophages from mouse bone marrow cells

Macrophages play essential roles in tissues, wherein they exert beneficial or detrimental functions depending on the signals they encounter during their differentiation. Here, we present a protocol to differentiate mouse bone marrow cells into macrophages under specific cues to evaluate their impact on macrophage phenotypic acquisition. We provide detailed instructions for optimal cell culture conditions, quality controls, and examples of concluding immunological functional assays. This protocol is applicable in short- and long-term drug-based modulation of macrophage functions. For complete details on the use and execution of this protocol, please refer to Scafidi et al.¹.