Abstract
Macrophages play a crucial role in mediating antibody-dependent cellular phagocytosis (ADCP), a process that enhances the effectiveness of several cancer immunotherapies. In this protocol, we outline detailed steps for isolating, differentiating, and polarizing macrophages from both mouse and human sources. Additionally, we describe an optimized technique to assess ADCP using in vitro co-culture studies followed by flow cytometry analysis. This protocol offers a reliable approach to evaluate the efficiency of macrophage-mediated phagocytosis of cancer cells during treatment with antibody-based therapies. For complete details on the use and execution of this protocol, please refer to Osorio et al.1.