Abstract
Osteoclasts are the only cells that are capable of resorbing bones, and they are involved in multiple diseases and disorders. This chapter will describe several in vitro osteoclastogenesis methods, which allows further investigation of molecular mechanisms of osteoclastogenesis in normal physiological and disease conditions. This chapter includes a protocol for isolating osteoclast progenitors from mouse bone marrow and human peripheral blood, as well as obtaining murine osteoblasts for the coculture system. Furthermore, culture and identification of multinucleated osteoclasts in vitro is also described in this chapter.
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References
Teitelbaum SL (2000) Bone resorption by osteoclasts. Science 289(5484):1504–1508
Manolagas SC (2000) Birth and death of bone cells: basic regulatory mechanisms and implications for the pathogenesis and treatment of osteoporosis. Endocr Rev 21(2):115–137
Goldring SR, Gravallese EM (2000) Pathogenesis of bone erosions in rheumatoid arthritis. Curr Opin Rheumatol 12(3):195–199
Olechnowicz SW, Edwards CM (2014) Contributions of the host microenvironment to cancer-induced bone disease. Cancer Res 74(6):1625–1631
Morand EF, Leech M, Bernhagen J (2006) MIF: a new cytokine link between rheumatoid arthritis and atherosclerosis. Nat Rev Drug Discov 5(5):399–410
Santos LL et al (2008) Reduced arthritis in MIF deficient mice is associated with reduced T cell activation: down-regulation of ERK MAP kinase phosphorylation. Clin Exp Immunol 152(2):372–380
Herrero LJ et al (2011) Critical role for macrophage migration inhibitory factor (MIF) in Ross River virus-induced arthritis and myositis. Proc Natl Acad Sci U S A 108(29):12048–12053
Gu R et al (2015) Macrophage migration inhibitory factor is essential for osteoclastogenic mechanisms in vitro and in vivo mouse model of arthritis. Cytokine 72(2):135–145
Suda T et al (1999) Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr Rev 20(3):345–357
Arai F et al (1999) Commitment and differentiation of osteoclast precursor cells by the sequential expression of c-Fms and receptor activator of nuclear factor kappaB (RANK) receptors. J Exp Med 190(12):1741–1754
Kim JH, Kim N (2016) Signaling pathways in osteoclast differentiation. Chonnam Med J 52(1):12–17
Eeles DG et al (2015) Osteoclast formation elicited by interleukin-33 stimulation is dependent upon the type of osteoclast progenitor. Mol Cell Endocrinol 399:259–266
Lakkakorpi PT, Vaananen HK (1996) Cytoskeletal changes in osteoclasts during the resorption cycle. Microsc Res Tech 33(2):171–181
Vaananen HK et al (2000) The cell biology of osteoclast function. J Cell Sci 113(Pt 3):377–381
Bradley EW, Oursler MJ (2008) Osteoclast culture and resorption assays. Methods Mol Biol 455:19–35
Wu DJ et al (2014) A novel in vivo gene transfer technique and in vitro cell based assays for the study of bone loss in musculoskeletal disorders. J Vis Exp 88. https://doi.org/10.3791/51810
Allan EH et al (2003) Differentiation potential of a mouse bone marrow stromal cell line. J Cell Biochem 90(1):158–169
Al-Abed Y, VanPatten S (2011) MIF as a disease target: ISO-1 as a proof-of-concept therapeutic. Future Med Chem 3(1):45–63
Chang MK et al (2008) Osteal tissue macrophages are intercalated throughout human and mouse bone lining tissues and regulate osteoblast function in vitro and in vivo. J Immunol 181(2):1232–1244
Singh PP et al (2012) Membrane-bound receptor activator of NFkappaB ligand (RANKL) activity displayed by osteoblasts is differentially regulated by osteolytic factors. Biochem Biophys Res Commun 422(1):48–53
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Gu, R. (2020). Methods to Determine the Effects of MIF on In Vitro Osteoclastogenesis Using Murine Bone Marrow-Derived Cells and Human Peripheral Blood Mononuclear Cells. In: Harris, J., Morand, E. (eds) Macrophage Migration Inhibitory Factor. Methods in Molecular Biology, vol 2080. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9936-1_12
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DOI: https://doi.org/10.1007/978-1-4939-9936-1_12
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