21B. Standardization of the Differentiation of THP-1 Monocytes into Macrophages

Lead Presenter(s)

Abigail Allen

Abstract or Description

Macrophages play a crucial role in the immune system, defending against pathogens and facilitating tissue repair following infection. Derived from monocytes, the lifecycle and functions of macrophages are central to many research studies on both health and diseases. Macrophages can be differentiated into an inflammatory phenotype (M1) or an anti-inflammatory phenotype (M2) depending on cytokine exposure. The THP-1 monocyte cell line, an immortalized source of monocytes, is widely used to study macrophage differentiation in vitro because they provide an abundance of cells for downstream applications and a uniform source of macrophages when compared with primary cells. However, the differentiation protocol for converting THP-1 monocytes into macrophages is highly variable across different studies. This research aims to standardize the differentiation process, developing a reliable and reproducible protocol. Herein, we investigated the effects of varying concentrations and exposure times of PMA, stimuli (IL-4, LPS, IFN-γ) and rest periods prior to stimulation. RTPCR was used to assess macrophage differentiation and M1/M2 skewing. Our findings demonstrate that PMA at a concentration between 5-20ng/mL effectively differentiates THP-1 monocytes into macrophages. In the future, we plan to further explore the skewing of M1 and M2 macrophages through chemokine exposure and rest period variations.

Mentor

Timothy Sparer