Osteoclast Differentiation Assay

Methods Mol Biol. 2019:1882:143-148. doi: 10.1007/978-1-4939-8879-2_12.

Abstract

Osteoclasts are highly specialized multinucleated cells derived from the monocyte/macrophage hematopoietic lineage that are uniquely capable of adhering to bone matrix and resorbing bone. The tartrate-resistant acid phosphatase (TRAP) assay is the most common method to detect osteoclasts population in vitro. Here we described a general protocol of inducing osteoclast differentiation from the murine macrophage cell line, RAW264.7, and identification of osteoclasts with the classical TRAP assay.

Keywords: Bone; MCSF; Osteoblasts; RANK; RANKL; RAW264.7; TRAP staining.

MeSH terms

  • Animals
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods*
  • Cell Differentiation*
  • Macrophage Colony-Stimulating Factor / metabolism
  • Mice
  • Osteoclasts / physiology*
  • RANK Ligand / metabolism
  • RAW 264.7 Cells
  • Recombinant Proteins / metabolism
  • Tartrate-Resistant Acid Phosphatase / analysis*

Substances

  • CSF1 protein, human
  • RANK Ligand
  • Recombinant Proteins
  • TNFSF11 protein, human
  • Macrophage Colony-Stimulating Factor
  • Tartrate-Resistant Acid Phosphatase