Murine osteoclasts were prepared from bone marrow cells as previously described (Kim et al., 2007 (link); 2015 (link)). In brief, mouse bone marrow cells were isolated from tibiae and femurs of 6–8 week old ICR mice by flushing the bone marrows with α-minimal essential medium (α-MEM). Bone marrow cells were cultured in α-MEM (Hyclone Laboratories, USA), containing 10% fetal bovine serum (FBS) with M-CSF (30 ng/ml), for 3 days. Adherent cells [bone marrow-derived macrophage-like cells (BMMs)] were further cultured with M-CSF (30 ng/ml) and RANKL (20–150 ng/ml) for 3 days. Cultured cells were fixed and stained for TRAP. TRAP-positive cells with more than three nuclei were counted as osteoclasts.
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Murine Osteoclast Differentiation Protocol
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Bone marrow cells
Bone marrow derived macrophage
Bone marrows
Cells
Cultured cells
Cultured medium
Femurs
Fetal bovine serum
Icr mice
M csf
Mouse
Nuclei
Osteoclasts
Rankl
Tibiae
Corresponding Organization : Chonnam National University
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Variable analysis
independent variables
- M-CSF (30 ng/ml)
- RANKL (20–150 ng/ml)
- TRAP-positive cells with more than three nuclei (counted as osteoclasts)
- Mouse bone marrow cells isolated from tibiae and femurs of 6–8 week old ICR mice
- Bone marrow cells cultured in α-MEM (Hyclone Laboratories, USA), containing 10% fetal bovine serum (FBS)
- Adherent cells [bone marrow-derived macrophage-like cells (BMMs)] further cultured with M-CSF (30 ng/ml) and RANKL (20–150 ng/ml) for 3 days
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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