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Updated: Aug 10, 2026
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Osteoclast Derivation from Mouse Bone Marrow
Published on: November 6, 2014
Enrichment of generated murine osteoclasts
A Shioi1, F P Ross, S L Teitelbaum
1Department of Pathology and Laboratory Medicine, Jewish Hospital at Washington University Medical Center, St. Louis, MO 63110.
Calcified Tissue International
|November 1, 1994
Summary
Researchers developed a method to isolate pure, viable murine osteoclasts from bone marrow cultures. This technique utilizes collagenase treatment to remove stromal cells, enabling further study of osteoclast biology.
Area of Science:
- Cell Biology
- Skeletal Biology
- Immunology
Background:
- Osteoclast purity is crucial for biochemical and molecular studies.
- Avian osteoclasts are easily generated in vitro from pure precursors.
- Murine osteoclast propagation is inefficient and requires stromal cells.
Purpose of the Study:
- To develop a method for enriching murine osteoclasts in culture while maintaining viability.
- To investigate the mechanisms by which stromal cells promote murine osteoclast generation and survival.
Main Methods:
- Fractionated murine marrow cells were cultured with ST-2 stromal cells.
- Generated multinucleated giant cells were identified as osteoclasts by TRAP expression and bone resorption assays.
- Bacterial collagenase treatment was used to selectively remove stromal cells.
Main Results:
- A method yielding >60% pure, viable murine osteoclasts was developed.
- Isolated osteoclasts retained tartrate-resistant acid phosphatase (TRAP) expression and calcitonin responsiveness.
- Murine osteoclast generation requires contact with viable stromal cells and is promoted by secreted macrophage colony-stimulating factor (CSF-1).
Conclusions:
- A novel method for isolating pure murine osteoclasts was established.
- Stromal cell-derived CSF-1 is essential for murine osteoclastogenesis.
- This method facilitates further research into osteoclast function and regulation.
