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Establishment and characterization of factor-dependent macrophage cell lines
1Department of Microbiology, Saga Medical School, Japan.
Abstract:
Three macrophage cell lines from bone marrow cells of C3H/HeN mice were isolated by successive transfer of the cells in culture with L-cell-conditioned medium (LCM) or WEHI-3 cell-conditioned medium (WEHI-3CM). These cell lines, which express Fc receptors, are involved in Fc-mediated phagocytosis and possess nonspecific esterase activity. Two (BDM-1 and BDM-2) of three cell lines show dependency for growth on either macrophage colony-stimulating factor (M-CSF) (CSF-1) or granulocyte-macrophage colony-stimulating factor (GM-CSF) and do not respond to interleukin 3 (IL-3). The third clone (BDM-3) proliferates in response to IL-3 as well as to GM-CSF and weakly responds to M-CSF and to interleukin 4 (IL-4). GM-CSF, in combination with the suboptimal concentration of M-CSF, acted synergistically on the proliferation of BDM-1 cells. The tumor-promoting phorbol diester, 12-o-tetradecanoyl-phorbol-13-acetate (TPA) also acted synergistically with the three CSFs (IL-3, GM-CSF, and M-CSF) to stimulate the proliferation of BDM-1 cells. The synergistic effect was observed when cells were pretreated with TPA and subsequently stimulated with IL-3. The calcium ionophore A23187 enhanced the proliferation of BDM-1 cells costimulated with TPA and IL-3. These factor-dependent macrophage cell lines should be useful for studying signal transduction mechanisms in the regulation of cell growth.
Insights
New macrophage cell lines were developed from mouse bone marrow. These factor-dependent cell lines, expressing Fc receptors, are valuable tools for investigating macrophage growth regulation and signal transduction pathways.
Area of Science:
- Immunology
- Cell Biology
- Hematopoiesis
Background:
- Macrophage cell lines are crucial for studying immune responses and cellular processes.
- Understanding the regulation of macrophage proliferation is essential for immunology and disease research.
Purpose of the Study:
- To isolate and characterize novel, factor-dependent macrophage cell lines from mouse bone marrow.
- To investigate the growth factor requirements and responses of these newly established cell lines.
Main Methods:
- Isolation of macrophage cell lines (BDM-1, BDM-2, BDM-3) from C3H/HeN mouse bone marrow.
- Culture with L-cell-conditioned medium (LCM) or WEHI-3 cell-conditioned medium (WEHI-3CM).
- Assays for Fc receptor expression, Fc-mediated phagocytosis, esterase activity, and proliferation in response to various cytokines (M-CSF, GM-CSF, IL-3, IL-4) and signaling molecules (TPA, A23187).
Main Results:
- Three distinct macrophage cell lines were established, exhibiting Fc receptors and phagocytic activity.
- BDM-1 and BDM-2 are dependent on M-CSF or GM-CSF, while BDM-3 responds to IL-3 and GM-CSF.
- Synergistic proliferation effects were observed with combinations of growth factors (GM-CSF + M-CSF) and with TPA (tumor promoter) plus cytokines, enhanced by calcium ionophore A23187.
Conclusions:
- The developed factor-dependent macrophage cell lines provide valuable models for studying macrophage biology.
- These cell lines are useful for dissecting signal transduction pathways regulating macrophage proliferation and function.
- The findings highlight complex regulatory mechanisms involving cytokines, TPA, and calcium signaling in macrophage growth.