ArticlePloS one2022
Evaluation of culture conditions for osteoclastogenesis in RAW264.7 cells.
Article in PloS one, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 23 citations in OpenAlex.
- Microbial-Derived Daidzin (Eco-3) Suppresses RANKL-Induced Osteoclastogenesis by Targeting the TBK1-NFATc1 Signaling Axis.International journal of molecular sciences · 2026Article
- Human parvovirus B19 VP1u‑induced osteoclastogenesis is enhanced by RANKL in RAW264.7 macrophage cells.Molecular medicine reports · 2026Article
- Hydrogen Sulfide Suppresses RANKL-Induced Osteoclast Differentiation by Modulating Nrf2/GPX4 Signaling.Physiological research · 2026Article
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- Bridging the gap in OA therapeutics: Bioengineered strategies to target osteoclast-chondrocyte crosstalk.Bioengineering & translational medicine · 2026Review
- Bone sialoprotein: a multifunctional regulator of bone remodelling and tumour progression.Bone research · 2026Review
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- OptimizingPeerJ · 2026Article
- Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells.Materials today. Bio · 2025Article
- Binary curcuminoid complex CRE-Bin inhibits osteoclast differentiation by suppressing the canonical NF-κB signaling pathway and modulating microRNA-223 expression.Scientific reports · 2025Article
- Guidance on the assessment of the functionality of biomaterials for periodontal tissue regeneration: Methodologies and testing procedures.Dental materials : official publication of the Academy of Dental Materials · 2025Review
- Targeting dipeptidyl peptidase-8/9 to combat inflammation-induced osteoclastogenesis in RAW264.7 macrophages and analysis of anti-osteoclastogenesis potential of chrysin.Iranian journal of basic medical sciences · 2025Article
- C-X-C motif chemokine ligand 13 suppresses osteoclast differentiation via interference with RANKL-RANK interaction.Frontiers in immunology · 2025Article
- Exploring theMetabolites · 2024Article
- A novel method to efficiently differentiate human osteoclasts from blood-derived monocytes.Biological procedures online · 2024Article
- In Vitro Effects ofJournal of functional biomaterials · 2024Article
- Extracellular Signal-Regulated Kinases Play Essential but Contrasting Roles in Osteoclast Differentiation.International journal of molecular sciences · 2023Review
- Modified Curcuminoid-Rich Extract Liposomal CRE-SDInhibits Osteoclastogenesis via the Canonical NF-κB Signaling Pathway.Pharmaceutics · 2023Article
Corrections and comments
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Osteoclasts are the only multinucleated cells in vivo responsible for bone resorption and are vital for regulating bone remodeling and maintaining bone mass. The RAW264.7 cell line is widely used to study osteoclastic differentiation and biological molecular mechanism. However, protocols for inducing osteoclast formation in RAW264.7 cells vary considerably between laboratories, hindering the replication of results. Therefore, we tested the influence of culture conditions on osteoclast differentiation, including cell density and receptor activator of nuclear factor kappa-B ligand (RANKL) concentrations with or without macrophage colony-stimulating factors (M-CSF). Tartrate-resistant acid phosphatase (TRAP) staining was used to detect the morphology of osteoclasts. qPCR was used to detect gene expression of osteoclast-specific gene marker cathepsin K (CTSK), osteoclast transcription factors c-Fos and nuclear factor of activated T cells, cytoplasmic 1 (NFATc1). The bone resorption function was evaluated by a scanning electron microscope (SEM). RANKL treatment increased multinucleated osteoclasts formation and increased CTSK, c-Fos and NFATc1 gene expression. Compared with RANKL treatment, M-CSF significantly decreased multinucleated osteoclasts formation, reduced CTSK gene expression and had little effect on c-Fos and NFATc1 gene expression. Concerning bone resorption activity, RANKL treatment increased bone resorption pits on bovine bone slices. Significantly higher levels of osteoclastogenesis were observed with RAW264.7-cell density of 2×104 cells/well in 24-well plates. Our results suggest that the addition of 50 ng/ml M-CSF has no positive effect on osteoclastogenesis. RANKL treatment and cell density contribute to osteoclast formation, and the optimal conditions are beneficial when exploring osteoclast function and mechanism.
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