JoVE Encyclopedia of Experiments
Immunology
0 views • 4:27 min • July 8th, 2025
Osteoclastogenesis is the process by which osteoclast precursors differentiate and fuse to form multinucleated osteoclasts.
To study the effect of the anti-c-fms antibody on osteoclastogenesis in vitro, take a culture plate containing a mouse bone marrow cell suspension. Add macrophage colony-stimulating factor, M-CSF.
In culture, M-CSF binds to its specific receptor, c-fms, on myeloid precursor cells, activating signaling pathways and causing differentiation into bone marrow macrophages, BMMs, which represent osteoclast precursors and adhere to the plate.
Remove the non-adherent cells. Harvest the BMMs, and seed them onto multi-well plate wells. Supplement with M-CSF and receptor activator of nuclear factor kappa-Β ligand, RANKL.
Add increasing anti-c-fms antibody concentrations to the wells.
In the control well, M-CSF binds to the c-fms receptors and activates signaling pathways, stimulating RANK receptor expression and promoting BMM survival and proliferation. RANKL binds to the RANK receptors, triggering a signaling cascade and causing BMM differentiation and fusion into osteoclasts.
In wells with higher antibody concentrations, the antibodies bind and effectively block the c-
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