In Vitro Screening Method for Characterization of Macrophage Activation Responses

Methods Protoc. 2022 Aug 30;5(5):68. doi: 10.3390/mps5050068.

Abstract

Macrophage activation refers to the enhanced functionality of macrophages in response to endogenous or exogenous stimuli. Due to the existence of limitless stimuli and a multitude of receptors on macrophage surfaces, the nature of activation (or acquired functioning) can be specific to the encountering stimulus. This article describes a macrophage-activation screening platform in a 96-well format. The methodology involves the generation of bone marrow-derived macrophages, their activation into two extreme activation states, and screening of activated macrophages for expression of bonafide protein biomarkers. A high-throughput and stringent assay to determine macrophage activation markers developed in this article can be adapted for biomarker determination in pathological conditions and toxicant/drug safety screening.

Keywords: BMDM; M1; M2; alternatively-activated macrophages (AAM); classically-activated macrophages (CAM); macrophage; macrophage activation.