Establishment and characterization of macrophage-like cell lines expressing osteoclast-specific markers

J Bone Miner Metab. 2001;19(4):220-7. doi: 10.1007/s007740170024.

Abstract

Osteoclasts differentiate from hematopoietic precursors of the monocyte/macrophage lineage to mononuclear preosteoclasts and multinuclear mature osteoclasts. In the present study, we report on the establishment of macrophage like cell lines from mouse bone marrow by coculturing bone marrow cells with mouse chondrocytes. Isolated clones (MLC-6 and MLC-7 cells) expressed fully differentiated osteoclast markers, such as calcitonin receptors, vitronectin receptors, tartrate-resistant acid phosphatase and vacuolar H+ -ATPase, in the absence of osteoclast differentiation factor/osteoprotegerin ligand/RANKL/TRANCE, which was essential for osteoclast differentiation. Most clones also maintained expression of a macrophage-associated protein, namely non-specific esterase. Both MLC-6 and MLC-7 cells released cathepsin K into the culture medium. Both clones resolved dentine slices when cocultured with the osteoblast cell line ST2. The cloned cell lines are considered to be useful tools in the study of osteoclast differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins*
  • Base Sequence
  • Biomarkers
  • Cell Differentiation
  • Cell Line
  • Coculture Techniques
  • DNA Primers / genetics
  • Gene Expression
  • Glycoproteins / genetics
  • Immunohistochemistry
  • Macrophages / cytology*
  • Macrophages / metabolism*
  • Matrix Metalloproteinase 9 / genetics
  • Mice
  • Microscopy, Electron, Scanning
  • Osteoclasts / cytology*
  • Osteoclasts / metabolism*
  • Osteoprotegerin
  • RNA-Binding Proteins / genetics
  • Receptors, Calcitonin / genetics
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Tumor Necrosis Factor
  • Receptors, Vitronectin / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factors / genetics
  • Vacuolar Proton-Translocating ATPases / metabolism

Substances

  • Bacterial Proteins
  • Biomarkers
  • DNA Primers
  • Glycoproteins
  • MtrB protein, Bacteria
  • Osteoprotegerin
  • RNA-Binding Proteins
  • Receptors, Calcitonin
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Tumor Necrosis Factor
  • Receptors, Vitronectin
  • Tnfrsf11b protein, mouse
  • Transcription Factors
  • Matrix Metalloproteinase 9
  • Vacuolar Proton-Translocating ATPases