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Updated: Aug 19, 2026

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
Published on: March 15, 2018
Macrophage colony-stimulating factor and interleukin-1 alpha maintain the survival of osteoclast-like cells
1Department of Biochemistry, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Abstract:
When mouse bone marrow cells were co-cultured with a stromal cell line. ST2, numerous osteoclast-like multinucleated cells (MNCs) were formed. To enrich the MNCs which were tartrate-resistant acid phosphatase (TRAP)-positive, we treated the cultures with dispase. Enriched TRAP-positive MNCs adhering to the bottom of the dish were cultured for a further 24h with medium containing 15% fetal calf serum. More than 80% of TRAP-positive MNCs were detached from the dish during the culture. However, when ST2 cells were included during the culture period, survival of TRAP-positive MNCs were significantly increased. Moreover, among various factors examined for the survival of TRAP-positive MNCs, only M-CSF and IL-1 alpha were effective in prolonging viability of TRAP-positive MNCs.
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