Generation of osteoclasts in vitro, and assay of osteoclast activity.
Osteoclasts are bone-resorbing multinucleated cells derived from the monocyte-macrophage lineage. The authors have developed a mouse marrow culture system and a coculture system of mouse osteoblasts and hemopoietic cells, in which osteoclasts are formed in response to various osteotropic factors such as 1alpha,25-dihydroxyvitamin D3, parathyroid hormone, prostaglandin E2, and interleukin-11. Recent studies have revealed that osteoblasts express two cytokines essential for osteoclastogenesis: receptor activator of nuclear factor kappaB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). Using RANKL and M-CSF, we can induce osteoclasts from monocyte-macrophage lineage cells even in the absence of osteoblasts. This chapter describes the methods for osteoclast formation in vitro in the presence and absence of osteoblasts, and for pit-formation assay using dentine slices and osteoclasts formed in vitro. These culture systems have made it possible to investigate each step of osteoclast development and function separately.
Takahashi
Naoyuki
N
Udagawa
Nobuyuki
N
Kobayashi
Yasuhiro
Y
Suda
Tatsuo
T
D000199:Actins / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D001862:Bone Resorption / Q000378:metabolism / Q000473:pathology; D002021:Buffers; D056657:Cathepsin K; D002403:Cathepsins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D018920:Coculture Techniques / Q000379:methods; D003094:Collagen; D003470:Culture Media; D005782:Gels; D007150:Immunohistochemistry / Q000379:methods; D016173:Macrophage Colony-Stimulating Factor / Q000378:metabolism / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology; D051379:Mice; D008954:Models, Biological; D010006:Osteoblasts / Q000166:cytology; D010010:Osteoclasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D053245:RANK Ligand / Q000378:metabolism / Q000494:pharmacology
D000199:Actins; D002021:Buffers; D003470:Culture Media; D005782:Gels; D053245:RANK Ligand; C505689:Tnfsf11 protein, mouse; D016173:Macrophage Colony-Stimulating Factor; D003094:Collagen; D002403:Cathepsins; D056657:Cathepsin K; C534132:Ctsk protein, mouse