Bone marrow-derived macrophage

Bone-marrow-derived macrophage (BMDM) refers to macrophage cells that are generated in a research laboratory from mammalian bone marrow cells.[1][2][3] BMDMs can differentiate into mature macrophages in the presence of growth factors and other signaling molecules.[1][2] Undifferentiated bone marrow cells are cultured in the presence of macrophage colony-stimulating factor (M-CSF; CSF-1).[3] M-CSF is a cytokine and growth factor that is responsible for the proliferation and commitment of myeloid progenitors into monocytes (which then mature into macrophages).[3][4] Macrophages have a wide variety of functions in the body including phagocytosis of foreign invaders and other cellular debris, releasing cytokines to trigger immune responses, and antigen presentation.[2] BMDMs provide a large homogenous population of macrophages that play an increasingly important role in making macrophage-related research possible and financially feasible.[5]

Production
Schema of in vitro BMDM production

In order to produce BMDMs, mesenchymal stem cells are removed from the tibia or femur of mice.[6] Since BMDMs are derived from bone marrow, withdrawn cells are healthy and naïve (or unactivated), regardless of the condition of donor mice.[5] After removal, stem-cells are incubated with CSF-1.[6] Without CSF-1, the cells enter an inactive state but can reinitiate growth and differentiation if stimulated later.[6] Mature macrophages and fibroblasts, which may carry unwanted growth factors, are removed.[6] Next, IL-3 and IL-1, two growth factors, are often added to increase yield and promote rapid terminal differentiation.[6] Exogenous media containing growth factors and other serums must also be added to make the cells continually viable.[6] Full growth and differentiation take approximately 5–8 days.[6]

Millions of BMDMs can be derived from one mouse and frozen for years. After being thawed BMDMs can respond to a variety of stimuli such as LPS, IFN-γ, PAMPs, NF-κB, and IRF3.[1][5][7] These signals induce translation of genes that produce cytokines and determine if macrophages are M1 (pro-inflammatory) or M2 (anti-inflammatory).[2] If BMDMs are not frozen, they age and become less viable as CSF-1 and growth factors in their media decreases.[1]

Proliferation of BMDMs can also be inhibited by a number of reagents.[6] For example, growth and differentiation is dependent on CSF-1 and a functional CSF-1 receptor, a member of the tyrosine kinase family.[6] Without a functional CSF-1 receptors, stem cells cannot respond to CSF-1 stimuli and therefore cannot differentiate; interferons can cause a down regulation of the CSF-1 receptor.[6] Additionally, without stimuli like LPS to induce macrophage maturation to M1 or M2, mice accumulate a large pool of monocytes, the precursor cells to macrophages, which are less helpful for macrophage-specific research[6]

References
  1. Barrett JP, Costello DA, O'Sullivan J, Cowley TR, Lynch MA (April 2015). "Bone marrow-derived macrophages from aged rats are more responsive to inflammatory stimuli". Journal of Neuroinflammation. 12 (1): 67. doi:10.1186/s12974-015-0287-7. PMC 4397943. PMID 25890218.
  2. Li, Yue; Niu, Shixian; Xi, Dalin; Zhao, Shuqi; Sun, Jiang; Jiang, Yong; Liu, Jinghua (2019). "Differences in Lipopolysaccharides-Induced Inflammatory Response Between Mouse Embryonic Fibroblasts and Bone Marrow-Derived Macrophages". Journal of Interferon & Cytokine Research. 39 (6): 375–382. doi:10.1089/jir.2018.0167. ISSN 1557-7465. PMID 30990360.
  3. Weischenfeldt J, Porse B (December 2008). "Bone Marrow-Derived Macrophages (BMM): Isolation and Applications". Cold Spring Harbor Protocols. 2008 (12): pdb.prot5080. doi:10.1101/pdb.prot5080. PMID 21356739.
  4. Hamilton, Thomas A.; Zhao, Chenyang; Pavicic, Paul G.; Datta, Shyamasree (2014-11-21). "Myeloid Colony-Stimulating Factors as Regulators of Macrophage Polarization". Frontiers in Immunology. 5: 554. doi:10.3389/fimmu.2014.00554. ISSN 1664-3224. PMC 4240161. PMID 25484881.
  5. Marim, Fernanda M.; Silveira, Tatiana N.; Lima, Djalma S.; Zamboni, Dario S. (2010-12-17). "A method for generation of bone marrow-derived macrophages from cryopreserved mouse bone marrow cells". PLOS ONE. 5 (12): e15263. Bibcode:2010PLoSO...515263M. doi:10.1371/journal.pone.0015263. ISSN 1932-6203. PMC 3003694. PMID 21179419.
  6. Hume, D. A.; Allan, W.; Fabrus, B.; Weidemann, M. J.; Hapel, A. J.; Bartelmez, S. (1987). "Regulation of proliferation of bone marrow-derived macrophages". Lymphokine Research. 6 (2): 127–139. ISSN 0277-6766. PMID 3035291.
  7. Oppong-Nonterah, Gertrude O.; Lakhdari, Omar; Yamamura, Asami; Hoffman, Hal M.; Prince, Lawrence S. (2019). "TLR Activation Alters Bone Marrow-Derived Macrophage Differentiation". Journal of Innate Immunity. 11 (1): 99–108. doi:10.1159/000494070. ISSN 1662-8128. PMC 6296861. PMID 30408777.
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