Effect of the Anti C-fms Antibody on Osteoclastogenesis in Mouse Bone Marrow Cells In Vitro

Published: August 31, 2023

Abstract

Source: Marahleh, A., et al. Effect of Anti-c-fms Antibody on Osteoclast Formation and Proliferation of Osteoclast Precursor In Vitro. J. Vis. Exp. (2019)

In this video, we demonstrate the effect of different anti-c-fms antibody concentrations on osteoclastogenesis, which is the process of osteoclast formation from osteoclast progenitors.

Protocol

1. Generation of BMM Seed 1 x 107 cells/10 mL in a 10 cm culture dish and add M-CSF 100 ng/mL. The final concentration of M-CSF is 100 ng/mL of culture medium. Incubate the culture at 37 °C, 5% CO2 for 3 days. After 3 days, remove the culture medium, wash the cells vigorously with 10 mL of phosphate-buffered saline (PBS) twice to remove non-adherent cells, add 5 mL of room temperature 0.02% trypsin-EDTA in PBS, and incubate at 37 °C, 5% CO2…

Divulgations

The authors have nothing to disclose.

Materials

Anti-c-Fms antibody AFS98, a rat monoclonal, antimurine, c-Fms antibody (IgG2a)
RANKL PEPROTECH 315-11 Recombinant Murine sRANK Ligand, Source: E.coli
M-CSF Recombinant human M-CSF. 1/10 vol of CMG14–12 cell line culture supernatant at 5X106 cells in a 10-cm suspension culture dish.
α-MEM Wako with L-Glutamine and phenol red
Fetal Bovine Serum Biowest s1820-500 Fetal Bovine Serum French Origin
Culture dish Corning 100 mm x 20 mm style dish
96-well plate Thermofisher Scientific Nun clon Delta surface

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Effect of the Anti C-fms Antibody on Osteoclastogenesis in Mouse Bone Marrow Cells In Vitro. J. Vis. Exp. (Pending Publication), e21613, doi: (2023).

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