RRC ID 43150
Author Takahashi N, Udagawa N, Kobayashi Y, Suda T.
Title Generation of osteoclasts in vitro, and assay of osteoclast activity.
Journal Methods Mol Med
Abstract 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.
Volume 135
Pages 285-301
Published 2007-1-1
DOI 10.1007/978-1-59745-401-8_18
PII 1-59745-401-X:285
PMID 17951666
MeSH Actins / metabolism Animals Bone Marrow Cells / cytology Bone Resorption / metabolism Bone Resorption / pathology Buffers Cathepsin K Cathepsins / metabolism Cell Differentiation / drug effects Cell Line Cell Separation / methods Coculture Techniques / methods Collagen Culture Media Gels Immunohistochemistry / methods Macrophage Colony-Stimulating Factor / metabolism Macrophage Colony-Stimulating Factor / pharmacology Macrophages / cytology Mice Models, Biological Osteoblasts / cytology Osteoclasts / cytology* Osteoclasts / drug effects Osteoclasts / metabolism RANK Ligand / metabolism RANK Ligand / pharmacology
Resource
Human and Animal Cells RAW 264(RCB0535) ST2(RCB0224) MC3T3-G2/PA6(RCB1127)