Tran Thanh Thao *

* Correspondence: Tran Thanh Thao (email:

Main Article Content


Mouse bone marrow-derived macrophages (BMDM) have been identified as an important host cell model for studying mammalian macrophage functions during pathogen infection. Whereas colony stimulating factor (CSF)-1 is required for BMDM differentiation, the commercial CSF-1, however, is expensive. Therefore, L929 cell-conditioned medium (LCM) has been reported to be used as a source of CSF-1 in many recent studies. However, whether BMDM differentiated by commercial CSF-1 and LCM had any functional differences in bacterial infection, particularly Listeria monocytogenes (LM) infection, has remained unexplored. This study aimed to examine the morphology of macrophages differentiated from CSF-1 and LCM as well as the phagocytic function of these macrophages in LM infection. Mouse bone marrow cells were differentiated in CSF-1 or LCM through 6 days before infected by LM. Phagocytotic roles of BMDM were evaluated throug the capability of macrophages to take up the bacteria, as shown by infection assay and immunofluorescence microscope. The results showed that macrophages grown in CSF-1 and LCM were similar in morphology and phagocytic functions during LM infection. Macrophages from LCM-supplemented media were homogenous and obtained CD11b and F4/80 surface markers. Immunofluorescence images demonstrated that bone marrow-derived macrophages were able to successfully take up LM at 0.5 h but unable to control intracellular bacterial replication by 4 h post infection.
Keywords: bone marrow, bone marrow-derived macrophage, differentiation, infection, Listeria monocytogenes

Article Details


Becattini, S., Littmann, E. R., Carter, R. A. et al., 2017. Commensal microbes provide first line defense against Listeria monocytogenes infection. Journal of Experimental Medicine. 214(7): 1973-1989.

Birmingham, C. L., Canadien, V., Kaniuk, N. A., Steinberg, B. E., Higgins, D. E. and Brumell, J. H., 2008. Listeriolysin O allows Listeria monocytogenes replication in macrophage vacuoles. Nature. 451(7176): 350-354.

Gordon, S., 2007. The macrophage: past, present and future. European Journal of Immunology. 37(S1): S9-S17.

Henry, R., Shaughnessy, L., Loessner, M. J., Alberti‐Segui, C., Higgins, D. E. and Swanson, J. A., 2006. Cytolysin‐dependent delay of vacuole maturation in macrophages infected with Listeria monocytogenes. Cellular Microbiology. 8(1): 107-119.

Kuhn, M., Kathariou, S. and Goebel, W., 1988. Hemolysin supports survival but not entry of the intracellular bacterium Listeria monocytogenes. Infection and Immunity. 56(1): 79-82.

Lee, H.S., Stachelek, S.J., Tomczyk, N., Finley, M.J., Composto, R.J. and Eckmann, D.M., 2013. Correlating macrophage morphology and cytokine production resulting from biomaterial contact. Journal of Biomedical Materials Research. 101(1): 203-212.

Lin, H-S. and Gordon, S., 1978. Secretion of plasminogen activator by bone marrow-derived mononuclear phagocytes and its enhancement by colony-stimulating factor. Journal of Experimental Medicine. 150(2): 231-245.

Marim, F.M., Silveira, T.N., Lima, Jr. D.S. and Zamboni, D.S., 2010. A method for generation of bone marrow-derived macrophages from cryopreserved mouse bone marrow cells. PloS one. 5(12): e15263.

Mitchell, G., Ge, L., Huang, Q. et al., 2015. Avoidance of autophagy mediated by PlcA or ActA is required for Listeria monocytogenes growth in macrophages. Infection and Immunity. 83(5): 2175-2184.

Moore, K.J. and Matlashewski, G., 1994. Intracellular infection by Leishmania donovani inhibits macrophage apoptosis. The Journal of Immunology. 152(6): 2930-2937.

Myers, J.T., Tsang, A.W. and Swanson, J.A., 2003. Localized reactive oxygen and nitrogen intermediates inhibit escape of Listeria monocytogenes from vacuoles in activated macrophages. The Journal of Immunology. 171(10): 5447-5453.

Pyonteck, S.M., Akkari, L., Schuhmacher, A.J. et al., 2013. CSF-1R inhibition alters macrophage polarization and blocks glioma progression. Nature Medicine. 19(10): 1264.

Shen, A. and Higgins, D.E., 2005. The 5′ untranslated region‐mediated enhancement of intracellular listeriolysin O production is required for Listeria monocytogenes pathogenicity. Molecular Microbiology. 57(5): 1460-1473.

Stockinger, S., Materna, T., Stoiber, D. et al., 2002. Production of type I IFN sensitizes macrophages to cell death induced by Listeria monocytogenes. The Journal of Immunology. 169(11): 6522-6529.

Stuart, P.M., Cassell, G.H. and Woodward, J.G., 1990. Differential induction of bone marrow macrophage proliferation by mycoplasmas involves granulocyte-macrophage colony-stimulating factor. Infection and Immunity. 58(11): 3558-3563.

Taylor, P.R., Martinez-Pomares, L., Stacey, M., Lin, H-H., Brown, G.D. and Gordon, S., 2005. Macrophage receptors and immune recognition. Annual Review of Immunology. 23: 901-944.

Tilney, L.G. and Portnoy, D.A., 1989. Actin filaments and the growth, movement, and spread of the intracellular bacterial parasite, Listeria monocytogenes. The Journal of Cell Biology. 109(4): 1597-1608.

Weischenfeldt, J. and Porse, B., 2008. Bone marrow-derived macrophages (BMM): isolation and applications. Cold Spring Harbor Protocols. Accessed on 14 December 2018. Available from

Ying, W., Cheruku, P.S., Bazer, F.W., Safe, S.H. and Zhou, B., 2013. Investigation of macrophage polarization using bone marrow derived macrophages. Journal of Visualized Experiments. (76): e50323.