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…
開示
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 |
