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Isolation, Transfection, and Culture of Primary Human Monocytes
Published on: December 16, 2019
Isolation and Culturing of Primary Mouse and Human Macrophages
Alba de Juan1, Begoña Lavin Plaza2,3
1Institut Curie, PSL Research University, INSERM, U932, Paris, France.
Abstract:
Macrophages are mature immune cells involved in the defense of the organism. Since their discovery, the main function attributed to macrophages has been phagocytosis. However, in recent years, several new functions such as angiogenesis, tissue remodeling, clearance of apoptotic cells, pro- and anti-inflammatory properties and tumor growth have been attributed to macrophages. To perform such varied functions, macrophages acquire specific phenotypes in response to external signals. The possibility of replicating these phenotypes in vitro represents a cutting-edge tool to understand potential macrophage functions in vivo. This chapter outlines protocols used to isolate and culture murine bone marrow-derived and human monocyte-derived macrophages. Furthermore, macrophage polarization processes into different phenotypes, with special relevance to atherosclerosis, are indicated.
Insights
This study details methods for isolating and culturing macrophages, crucial immune cells. It explores macrophage polarization into distinct phenotypes, vital for understanding immune responses and diseases like atherosclerosis.
Area of Science:
- Immunology and Cell Biology
- Investigating the diverse roles and plasticity of macrophages in host defense and disease pathogenesis.
Background:
- Macrophages are key immune cells with diverse functions beyond phagocytosis, including angiogenesis and tissue remodeling.
- These functions are mediated by distinct macrophage phenotypes acquired in response to environmental cues.
- Understanding macrophage plasticity is crucial for deciphering their roles in health and disease.
Purpose of the Study:
- To provide protocols for in vitro isolation and culture of murine bone marrow-derived and human monocyte-derived macrophages.
- To describe the process of macrophage polarization into specific phenotypes.
- To highlight the relevance of macrophage polarization in the context of atherosclerosis.
Main Methods:
- Isolation of bone marrow cells from mice.
- Isolation of peripheral blood monocytes from humans.
- Culture and differentiation of monocytes into macrophages.
- Induction of macrophage polarization using specific stimuli.
- Characterization of macrophage phenotypes.
Main Results:
- Established protocols for generating bone marrow-derived macrophages (BMDMs) and monocyte-derived macrophages (MDMs).
- Demonstrated successful polarization of macrophages into distinct phenotypes in vitro.
- Highlighted phenotypic changes relevant to inflammatory conditions, particularly atherosclerosis.
Conclusions:
- In vitro culture and polarization of macrophages provide valuable models for studying their functions.
- Macrophage plasticity is a critical factor in immune responses and disease development.
- These protocols facilitate research into macrophage roles in atherosclerosis and other conditions.
