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Updated: May 15, 2026
Isolation and Culture of Bone Marrow-Derived Macrophages from Mice
Published on: June 23, 2023
In vitro derivation of macrophage from guinea pig bone marrow with human M-CSF
Karl O A Yu1, Steven A Porcelli, Howard A Shuman
1Section of Infectious Diseases, Department of Pediatrics, Comer Children's Hospital, University of Chicago Medical Center, Chicago, IL 60637, USA. karl.yu@uchospitals.edu
Abstract:
The guinea pig has a storied history as a model in the study of infectious disease and immunology. Because of reproducibility of data and availability of various reagents, inbred mice have since supplanted the guinea pig as the animal model-of-choice in these fields. However, several clinically-significant microorganisms do not cause the same pathology in mice, or mice may not be susceptible to these infections. These demonstrate the utility of other animal models - either as the primary method to study a particular infection, or to confirm or refute findings in the mouse before translating basic science into clinical practice. The mononuclear phagocyte, or macrophage (Mφ), plays a key role in antigen presentation and the pathogenesis of intracellular bacteria, such as Mycobacterium tuberculosis and Legionella pneumophila. Because of variable yield and difficult extraction from tissue, the preferred method of producing Mφ for in vitro studies is to expand murine bone marrow (BM) precursors with mouse macrophage colony-stimulating factor (M-CSF). This has not been shown in the guinea pig. Here, we report the empiric observation that human M-CSF - but not mouse M-CSF, nor human granulocyte/macrophage colony-stimulating factor - can be used to induce BM precursor differentiation into bonafide Mφ. The differentiated cells appeared as enlarged adherent cells, capable of both pinocytosis and large particle phagocytosis. Furthermore, we showed that these guinea pig BM-derived Mφ, similar to human monocyte/Mφ lines but unlike most murine BM Mφ, support growth of wild type L. pneumophila. This method may prove useful for in vitro studies of Mφ in the guinea pig, as well as in the translation of results found using mouse BM-derived Mφ towards studies in human immunology and infectious disease.
Insights
Human macrophage colony-stimulating factor (M-CSF) effectively differentiates guinea pig bone marrow precursors into macrophages. These macrophages support Legionella pneumophila growth, offering a new model for infectious disease and immunology research.
Area of Science:
- Immunology and Infectious Disease Research
- Animal Models in Biomedical Science
- Cell Biology and Cytokine Signaling
Background:
- Guinea pigs have historical significance in infectious disease and immunology research.
- Inbred mice are currently preferred but do not fully replicate all human infections.
- Macrophages play a critical role in host defense against intracellular pathogens like Legionella pneumophila.
Purpose of the Study:
- To establish a reliable method for generating guinea pig bone marrow-derived macrophages (Mφ) for in vitro studies.
- To investigate the utility of guinea pig Mφ in modeling infections that are not well-represented in mouse models.
- To compare the Mφ differentiation and pathogen support capabilities between guinea pig and mouse models.
Main Methods:
- Culturing guinea pig bone marrow precursors with various colony-stimulating factors (CSFs).
- Characterizing differentiated cells for macrophage morphology and function (pinocytosis, phagocytosis).
- Assessing the ability of guinea pig Mφ to support the growth of wild-type Legionella pneumophila.
Main Results:
- Human macrophage colony-stimulating factor (M-CSF) successfully induced differentiation of guinea pig bone marrow precursors into functional macrophages.
- These guinea pig Mφ exhibited typical macrophage characteristics, including adherence, pinocytosis, and phagocytosis.
- Unlike most murine Mφ, the guinea pig Mφ supported the growth of wild-type L. pneumophila, similar to human cell lines.
Conclusions:
- Human M-CSF provides a viable method for generating in vitro macrophage cultures from guinea pig bone marrow.
- Guinea pig-derived Mφ offer a valuable model for studying intracellular bacterial infections, particularly those poorly mimicked in mice.
- This approach facilitates research in guinea pig immunology and aids in translating mouse model findings to human studies.
