title: Cytokines synergistically induce osteoclast differentiation: support by immortalized or normal calvarial cells.
abstract: Conditionally immortalized murine calvarial (CIMC) cells that support differentiation of precursors into mature osteoclasts were isolated. All six CIMC cell lines supported osteoclast differentiation in response to 1,25-dihydroxyvitamin D(3) or interleukin (IL)-11. CIMC-4 cells also supported osteoclast differentiation in response to tumor necrosis factor (TNF)-alpha, IL-1beta, or IL-6. The resultant multinucleated cells expressed tartrate-resistant acid phosphatase and formed resorption lacunae on mineralized surfaces. CIMC-4 cells, therefore, establish an osteoclast differentiation assay that is responsive to many cytokines and does not rely on isolation of primary stromal support cells. Low concentrations of the cytokines synergistically stimulated differentiation when osteoclast precursors were cocultured with either CIMC-4 cells or primary calvarial cells. Osteoclast differentiation induced by all stimuli other than TNF-alpha was completely blocked by osteoprotegerin, whether the stimulators were examined alone or in combination. Moreover, study of precursors that lack TNF-alpha receptors showed that TNF-alpha induces osteoclast differentiation primarily through direct actions on osteoclast precursors, which is a distinct mechanism from that used by the other bone-resorptive agents examined in this study.
authors.affiliation: Department of Orthopaedics, Case Western Reserve University and University Hospitals of Cleveland, Ohio 44106-5000, USA.
authors.forename: Ashraf A
authors.initials: AA
authors.lastname: Ragab
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Nalepka
authors.forename: Yanming
authors.initials: Y
authors.lastname: Bi
authors.forename: Edward M
authors.initials: EM
authors.lastname: Greenfield
mesh_terms: D000135:Acid Phosphatase / Q000096:biosynthesis; D000818:Animals; D002117:Calcitriol / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005260:Female; D006023:Glycoproteins / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D017370:Interleukin-11 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D007527:Isoenzymes / Q000096:biosynthesis; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D010010:Osteoclasts / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D053244:Osteoprotegerin; D010281:Parathyroid Hormone / Q000494:pharmacology; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D012886:Skull / Q000166:cytology / Q000187:drug effects*; D013154:Spleen / Q000166:cytology; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*; D000071681:Tartrate-Resistant Acid Phosphatase; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D016207:Cytokines; D006023:Glycoproteins; D007375:Interleukin-1; D017370:Interleukin-11; D015850:Interleukin-6; D007527:Isoenzymes; D053244:Osteoprotegerin; D010281:Parathyroid Hormone; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; C506031:Tnfrsf11b protein, mouse; D014409:Tumor Necrosis Factor-alpha; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; D002117:Calcitriol