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Profiling of Macrophage Polarization Using Automated Enzyme-Linked Immunosorbent Assay
Simon Loevenich1, Vu To Nakstad1, Hanne Hein Trøen1
1Department of Biotechnology and Nanomedicine, SINTEF Industry, Trondheim, Norway.
Methods in Molecular Biology (Clifton, N.J.)
|September 30, 2025
Summary
This study details isolating human monocytes and polarizing them into M1 and M2 macrophage phenotypes. Cytokine levels are measured using enzyme-linked immunosorbent assays (ELISA) for immunology research.
Area of Science:
- Immunology
- Cell Biology
Background:
- Macrophage activation is crucial in immune responses.
- Macrophages polarize into distinct functional phenotypes (M1 and M2) based on environmental cues.
Purpose of the Study:
- To outline a protocol for isolating human primary CD14+ monocytes.
- To describe the polarization of these monocytes into M1 and M2 macrophage phenotypes.
Main Methods:
- Isolation of human primary CD14+ monocytes.
- Induction of M1 (pro-inflammatory) and M2 (anti-inflammatory) macrophage polarization.
- Detection of cytokine levels via manual and automated enzyme-linked immunosorbent assays (ELISA).
Main Results:
- Successful isolation of human CD14+ monocytes.
- Demonstrated polarization of monocytes into M1 and M2 phenotypes.
- Quantification of cytokine profiles associated with each phenotype using ELISA.
Conclusions:
- Provides a reproducible method for generating distinct human macrophage phenotypes in vitro.
- Enables the study of macrophage polarization in various immunological contexts.
- Facilitates the analysis of cytokine production for understanding immune signaling.
