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A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
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RANKL-Based Osteoclastogenic Assay from Murine Bone Marrow Cells
Zhenqiang Yao1, Lianping Xing1, Brendan F Boyce2
1Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, NY, USA.
Methods in Molecular Biology (Clifton, N.J.)
|November 16, 2020
Summary
Osteoclasts, crucial for bone health, can be generated in vitro from precursor cells. This method allows for the study of bone resorption and related diseases like osteoporosis.
Area of Science:
- Cell Biology
- Skeletal Biology
- Immunology
Background:
- Osteoclasts are essential for bone resorption, vital for skeletal development and maintaining bone homeostasis.
- Dysregulation of osteoclast activity leads to bone disorders such as osteopetrosis and osteoporosis.
- Osteoclasts originate from monocyte/macrophage lineage cells.
Purpose of the Study:
- To describe a protocol for generating osteoclasts in vitro.
- To outline methods for assessing osteoclast bone resorption activity.
Main Methods:
- Isolation of mouse bone marrow precursor cells.
- In vitro differentiation of osteoclasts using macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-κB ligand (RANKL).
- Assessment of osteoclast function by measuring resorption lacunae formation on bone slices.
Main Results:
- Successful generation of functional osteoclasts from mouse bone marrow cells.
- Demonstration of osteoclast resorptive activity through lacunae formation.
Conclusions:
- The described in vitro method provides a reliable system for studying osteoclast biology.
- This protocol facilitates research into bone diseases characterized by abnormal osteoclast activity.
