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• W2168094868 abstract "Osteoclasts (OC) are multinucleated bone resorbing cells that form via RANKL-induced fusion of heterogeneous mononuclear OC precursors (OCP). Currently, there are no unique surface markers to distinguish these OCP populations, which are diagnostic for erosive and metabolic bone diseases using culture assays. Thus, we investigated expression of DC-STAMP, a surface receptor required for OCP fusion, during osteoclastogenesis in vitro using a novel monoclonal antibody (1A2). Immunoprecipitation-Western blot analysis of OCP membrane proteins detected 106 kDa dimeric and 53 kDa monomeric DC-STAMP in non-denaturing and denaturing conditions, respectively, with greater sensitivity versus rabbit anti-sera (KR104). 1A2 also detected 99.9% of undifferentiated monocytes as a single population by flow cytometry with a MFI 100-fold over background, while KR104 was not useful in this assay. Functionally, 1A2 inhibited OCP fusion in vitro. RANKL stimulation of OCP induced DC-STAMPlo and DC-STAMPhi cells, which mature into OC and mononuclear cells respectively as determined by fluorescent microscopy and TRAP assays. Addition of DC-STAMPhi cells to purified DC-STAMPlo cultures produced larger, more nucleated OC vs. pure DC-STAMPlo cultures. RT-qPCR analysis of these two populations showed that OC markers (Trap and Oc-stamp) and fusogenic gene expression (Cd9 and Cd47), were significantly increased in DC-STAMPlo vs. DC-STAMPhi cells. Collectively, these results demonstrate that DC-STAMP is expressed on OCP as a dimer, which is efficiently detected by 1A2 via flow cytometry. RANKL induces osteoclastogenesis by stimulating DC-STAMP internalization in some OCP, and these DC-STAMPlo cells display the “master fusogen” phenotype. In contrast, DC-STAMPhi OCP can only act as mononuclear donors. J. Cell. Physiol. 223: 76–83, 2010. © 2009 Wiley-Liss, Inc." @default.
• W2168094868 created "2016-06-24" @default.
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• W2168094868 date "2009-01-01" @default.
• W2168094868 modified "2023-10-17" @default.
• W2168094868 title "RANKL induces heterogeneous DC-STAMP^loand DC-STAMP^hiosteoclast precursors of which the DC-STAMP^loprecursors are the master fusogens" @default.
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• W2168094868 doi "https://doi.org/10.1002/jcp.22012" @default.
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