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A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
Published on: March 15, 2018
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RANKL-based osteoclastogenic assays from murine bone marrow cells
Lianping Xing1, Brendan F Boyce1
1University of Rochester Medical Center, Rochester, NY, USA.
Methods in Molecular Biology (Clifton, N.J.)
|February 1, 2014
Summary
This protocol details generating osteoclasts, the bone-resorbing cells, from mouse bone marrow. This method is crucial for studying bone remodeling and diseases affecting the skeleton.
Area of Science:
- Cell Biology
- Skeletal Biology
- Immunology
Background:
- Osteoclasts are essential for bone resorption and remodeling.
- Dysfunctional osteoclasts contribute to skeletal pathologies.
- Mouse models are vital for studying osteoclast biology.
Purpose of the Study:
- To provide a standardized protocol for generating osteoclasts in vitro.
- To facilitate research on osteoclast function and bone diseases.
- To enable the study of genetic mouse models affecting bone metabolism.
Main Methods:
- Isolation of bone marrow cells from mice.
- Culture of cells using macrophage colony-stimulating factor (MCSF).
- Induction of osteoclast differentiation with receptor activator of NF-κB ligand (RANKL).
Main Results:
- Successful generation of functional osteoclasts from mouse bone marrow.
- Osteoclasts exhibited characteristic morphology and resorptive activity (data not shown).
- The protocol is reproducible and yields a sufficient number of cells for analysis.
Conclusions:
- This protocol offers a reliable method for in vitro osteoclast generation.
- It serves as a valuable tool for investigating bone remodeling and related disorders.
- Further studies can utilize these osteoclasts to explore therapeutic targets.
