Embryonic Stem Cells as a Model for Studying Osteoclast Lineage Development

  • Toshiyuki Yamane
  • Takahiro Kunisada
  • Shin-Ichi Hayashi
Part of the Methods in Molecular Biology™ book series (MIMB, volume 185)


Osteoclasts are cells specialized to resorb bone (1,2). They originate from hematopoietic progenitors (3,4), which are widely distributed throughout the body. Osteoclast progenitors differentiate into cells that are positive for calcitonin receptor and tartrate-resistant acid phosphatase (TRAP), which are specific markers for this lineage, and then they fuse with each other and form fully functional multinucleated large cells (1,2) (Fig. 1). TRAP-positive osteoclasts are found tightly attached to bone matrix (1,2). Two cytokines, macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-κB (RANK) ligand (RANKL; also known as OPGL, ODF, and TRANCE), are known to be responsible for this differentiation (5, 6, 7). In this chapter, we describe the procedure for the induction of differentiation into the osteoclast lineage from murine embryonic stem (ES) cells (8). We utilize co-culture systems with stromal cell lines, that is, bone marrow-derived ST2 cells (9) or newborn calvaria-derived M-CSF-deficient OP9 cells (10). M-CSF is produced constitutively by ST2, and the expression of RANKL is induced on ST2 by 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and dexamethasone (Dex) (6).
Fig. 1.

Scheme describing the development of osteoclast lineage. The essential molecules expressed in this lineage are indicated at the stage where maturation is arrested by the loss of function. The cells positive for TRAP are shaded.


Embryonic Stem Cell Osteoclast Progenitor Embryonic Stem Cell Culture Undifferentiated Embryonic Stem Cell Embryonic Stem Cell Culture Medium 
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Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Toshiyuki Yamane
    • 1
  • Takahiro Kunisada
    • 2
  • Shin-Ichi Hayashi
    • 1
  1. 1.Department of Immunology, School of Life Science, Faculty of MedicineTottori UniversityYonagoJapan
  2. 2.Department of Hygiene, Faculty of MedicineGifu UniversityGifuJapan

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