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Updated: Aug 22, 2026
Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages
Published on: April 20, 2018
Isolation and culture of murine macrophages
1Sir William Dunn School of Pathology, University of Oxford, UK.
Abstract:
The two most convenient sources of primary murine macrophages are the bone marrow and the peritoneal cavity. Resident peritoneal macrophages can readily be harvested from mice and purified by adherence to tissue culture plastic. The injection of Bio-Gel polyacrylamide beads or thioglycollate broth into the peritoneal cavity produces an inflammatory response allowing the purification of large numbers of elicited macrophages. The production of an activated macrophage population can be achieved by using Bacillus-Calmette-Guerin as the inflammatory stimulus. Resident bone marrow macrophages can be isolated following enzymatic separation of cells from bone marrow plugs and enrichment on 30% fetal calf serum containing medium or Ficoll-Hypaque gradients. Bone marrow-derived macrophages can be produced by differentiating nonadherent macrophage precursors with medium containing macrophage colony-stimulating factor.
Insights
Discover convenient methods for isolating primary murine macrophages from bone marrow and peritoneal sources. This summary covers techniques for obtaining resident, elicited, and activated macrophages for research applications.
Area of Science:
- Immunology
- Cell Biology
Background:
- Macrophages are crucial immune cells with diverse functions.
- Efficient isolation of primary murine macrophages is essential for immunological research.
Purpose of the Study:
- To outline accessible methods for obtaining primary murine macrophages.
- To describe techniques for isolating resident, elicited, and activated macrophage populations.
Main Methods:
- Harvesting resident peritoneal macrophages via adherence.
- Inducing inflammatory responses with Bio-Gel beads or thioglycollate for elicited macrophages.
- Utilizing Bacillus-Calmette-Guerin for activated macrophage production.
- Isolating bone marrow macrophages through enzymatic digestion and density gradient centrifugation.
- Differentiating bone marrow precursors using macrophage colony-stimulating factor.
Main Results:
- Established protocols for the isolation of various primary murine macrophage populations.
- Demonstrated the efficacy of different stimuli for macrophage elicitation and activation.
- Provided methods for both peritoneal and bone marrow-derived macrophage acquisition.
Conclusions:
- Bone marrow and peritoneal cavity offer convenient sources for primary murine macrophages.
- Specific protocols allow for the isolation of distinct macrophage populations (resident, elicited, activated) for experimental use.
