A Combination of Osteoclast Differentiation Factor and Macrophage-Colony Stimulating Factor Is Sufficient for both Human and Mouse Osteoclast Formation in Vitro

Abstract

Both human and murine osteoclasts can be derived in vitro from hematopoietic cells or monocytes that are co-cultured with osteoblasts or marrow-derived stromal cells. The osteoclastogenic stimulus provided by murine osteoblasts and marrow-derived stromal cells is now known to be mediated by osteoclast differentiation factor (ODF), a membrane-bound tumor necrosis factor-related ligand. This study demonstrates that mouse spleen cells and monocytes form osteoclasts when cultured in the presence of macrophage-colony stimulating factor (M-CSF) and a soluble form of murine ODF (sODF). Numerous multinucleated osteoclasts expressing tartrate resistant acid phosphatase (TRAP) and calcitonin receptor (CTR) formed within 7 days of culture and engaged in extensive lacunar bone resorption. Osteoclast number and bone resorption area was dependent on sODF concentration. Long-term cultured human monocytes also formed bone resorbing osteoclasts in response to co-stimulation by sODF and M-CSF, although this required more than 11 days in culture. This human osteoclast differentiation was strongly inhibited by granulocyte-macrophage colony stimulating factor. This study further characterises murine osteoclast differentiation caused by sODF and M-CSF co-stimulation in vitro, and shows that the same co-stimulation causes human osteoclast differentiation to occur. We propose that this methodology can be employed to investigate the direct effects of cytokines and other factors on human osteoclast differentiation.

RANKL:Osteoprotegerin ratio and bone mineral density in children with untreated juvenile dermatomyositis

“…One child had mild disease primarily affecting the skin, which resulted in misdiagnosis for 99.7 months. Patients had a mean Ϯ SD DAS of 11.1 Ϯ 3.5 (median 11, range [4][5][6][7][8][9][10][11][12][13][14][15][16][17]. Clinical data for the children with juvenile DM, including anthropometric measurements, are summarized in Table 2.…”

“…RANKL messenger RNA (mRNA) is expressed in trabecular bone and in lymphoid tissue active in mediating immune responses (14)(15)(16)(17). This protein expands the number of activated osteoclasts in the presence of macrophage colony-stimulating factor (14,15,17,18) and prevents apoptosis of mature osteoclasts (19). OPG is a soluble secreted decoy receptor of RANKL (20,21).…”

VEGF, FLT3 ligand, PlGF and HGF can substitute for M-CSF to induce human osteoclast formation: implications for giant cell tumour pathobiology

“…3,4 Osteoclast differentiation from monocyte or macrophage precursors requires the presence of receptor activator of nuclear factor kappa B ligand (RANKL) and macrophage-colony stimulating factor (M-CSF). 5 The importance of M-CSF in osteoclast formation is evidenced by the fact that in op/op osteopetrotic mice, which have a mutation in the M-CSF gene, very few osteoclasts are found in bone and there is markedly decreased bone resorption. 6,7 M-CSF promotes several aspects of osteoclastogenesis including the proliferation and fusion of osteoclast precursors as well as osteoclasts and the expression of the RANKL receptor by these cells.…”

Characterization of osteoclast precursors in human blood

“…OPGL, also known as TRANCE (TNF-related activation-induced cytokine) (Wong et al, 1997a), as RANKL (receptor activator of NF-kB ligand) (Anderson et al, 1997) and as ODF (osteoclast differentiation factor) , mediates the maturation of osteoclasts . OPG is a soluble receptor which inhibits OPGL-induced osteoclast formation and function in vivo and in vitro (Bucay et al, 1998;Fuller et al, 1998;Matsuzaki et al, 1998;Quinn et al, 1998;Burgess et al, 1999;. Overexpressing OPG transgenic mice (Simonet et al, 1997), as well as mice in which the OPGL gene is knocked out (Kong et al, 1999a), are osteopetrotic, whereas animals lacking OPG exhibit a high-turnover osteoporosis-like phenotype (Bucay et al, 1998).…”

“…The last two studies and those showing that multinucleated osteoclast-like cells develop from human peripheral blood mononuclear cells (PBMCs), when cultured with OPGL and CSF-1 (Matsuzaki et al, 1998;Quinn et al, 1998;, prompted the question of whether RANK is present on OCPs that circulate in peripheral blood (PB) ± without prior cytokine treatment. Early studies had demonstrated that OCPs originate from the haematopoietic stem cells (Sheven et al, 1986;Hagenaars et al, 1989;Hattersley & Chambers, 1989;Kurihara et al, 1989;Matayoshi et al, 1996) and circulate in PB (Walker, 1973(Walker, , 1975Kahn & Simmons, 1975;Coccia et al, 1980;Faust et al, 1999;.…”