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Updated: Jun 3, 2026
Isolation and Culture of Bone Marrow-Derived Macrophages from Mice
Published on: June 23, 2023
Murine bone marrow-derived macrophages
1Albert Einstein College of Medicine, Bronx, New York.
Abstract:
The molecular phagocytic lineage comprises, in order of increasing maturity, the committed macrophage precursor cell, the monoblast, promonocyte, monocyte and the macrophage. Methods for the preparation and culture of bone marrow-derived macrophages, developed by Stanley and colleagues (1-3), provide large numbers of mononuclear phagocytes that are capable of extensive cell proliferation. Since their proliferation can be stimulated by colony stimulating factor-1 (CSF-1), granulocyte macrophage colony stimulating factor (GM-CSF), or interleukin-3 (IL-3), they represent an important primary cell source for studies of the actions and interactions of these three growth factors. The principles underlying the method are: (1) to generate and expand primitive mononuclear phagocyte precursor cells by culturing bone marrow cells in a combination of partially purified CSF-1 and IL-3 for a period of 3 d, (2) to remove contaminating red cells, fibroblasts and mature macrophages and disrupt aggregates of proliferating cells, by proteolytic digestion of the nonadherent cells at d 1 and 3 of culture and, (3) to obtain a population of mononuclear phagocytes that is relatively homogeneous with respect to their state of differentiation by recovering only those cells (i.e., mono-blasts, promonocytes) that acquire the capacity to adhere to tissue culture plastic during d 4-5 of culture.
Insights
This study details a method for culturing bone marrow-derived macrophages, yielding pure populations of mononuclear phagocytes. These cells are crucial for investigating growth factors like colony-stimulating factor-1 (CSF-1) and interleukin-3 (IL-3).
Area of Science:
- Immunology
- Cell Biology
- Hematopoiesis
Background:
- The molecular phagocytic lineage includes precursor cells, monoblasts, promonocytes, monocytes, and macrophages.
- Stanley's method provides abundant mononuclear phagocytes for proliferation studies.
- These cells are stimulated by CSF-1, GM-CSF, and IL-3, making them vital for growth factor research.
Purpose of the Study:
- To present a method for preparing and culturing bone marrow-derived macrophages.
- To obtain a homogeneous population of mononuclear phagocytes for research.
- To facilitate studies on the actions and interactions of CSF-1, GM-CSF, and IL-3.
Main Methods:
- Culture bone marrow cells with CSF-1 and IL-3 to expand precursor cells.
- Use proteolytic digestion to remove non-proliferating cells and aggregates.
- Select cells that adhere to plastic between days 4-5 for homogeneity.
Main Results:
- Successful generation of large numbers of proliferating mononuclear phagocytes.
- Removal of contaminating cell types including red cells, fibroblasts, and mature macrophages.
- Isolation of a relatively homogeneous population of monoblasts and promonocytes.
Conclusions:
- The described method effectively produces pure, proliferating mononuclear phagocytes.
- This technique provides a valuable cell source for studying macrophage development and growth factor effects.
- The selective adherence property is key to achieving a homogeneous cell population.
