PMID- 27339271 OWN - NLM STAT- MEDLINE DCOM- 20180125 LR - 20180330 IS - 1549-4918 (Electronic) IS - 1066-5099 (Linking) VI - 34 IP - 11 DP - 2016 Nov TI - CXCR4(+) CD45(-) Cells are Niche Forming for Osteoclastogenesis via the SDF-1, CXCL7, and CX3CL1 Signaling Pathways in Bone Marrow. PG - 2733-2743 LID - 10.1002/stem.2440 [doi] AB - Bone homeostasis comprises the balance between bone-forming osteoblasts and bone-resorbing osteoclasts (OCs), with an acceleration of osteoclastic bone resorption leading to osteoporosis. OCs can be generated from bone marrow cells (BMCs) under the tightly regulated local bone environment. However, it remained difficult to identify the critical cells responsible for providing an osteoclastogenesis niche. In this study, we used a fluorescence-activated cell sorting technique to determine the cell populations important for forming an appropriate microenvironment for osteoclastogenesis and to verify the associated interactions between osteoclast precursor cells and non-OCs. We isolated and removed a small cell population specific for osteoclastogenesis (CXCR4(+) CD45(-) ) from mouse BMCs and cultured the remaining cells with receptor activator of nuclear factor-kappa B ligand (RANKL) and macrophage-colony stimulating factor. The resulting cultures showed significantly less large osteoclast formation. Quantitative RT-PCR analysis revealed that these CXCR4(+) CD45(-) cells expressed low levels of RANK and RANKL, but high levels of critical chemokines including stromal cell derived factor 1 (SDF-1), chemokine (C-X-C motif) ligand 7 (CXCL7), and chemokine (C-X3-C motif) ligand 1 (CX3CL1). Furthermore, an SDF-1-specific antibody strongly suppressed OC formation in RAW264.7 cells and antibodies against SDF-1, CXCL7, and CX3CL1 suppressed OC formation in BMCs. These results suggest that isolated CXCR4(+) CD45(-) cells support an appropriate microenvironment for osteoclastogenesis with a direct effect on the cells expressing SDF-1, CXCL7, and CX3CL1 receptors. The regulation of CXCR4(+) CD45(-) cell function might therefore inform therapeutic strategies for diseases involving loss of bone homeostasis. Stem Cells 2016;34:2733-2743. CI - (c) 2016 AlphaMed Press. FAU - Goto, Yoh AU - Goto Y AD - Department of Orthodontics, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan. AD - Department of Molecular Neurobiology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan. FAU - Aoyama, Mineyoshi AU - Aoyama M AD - Department of Molecular Neurobiology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan. AD - Department of Pathobiology, Nagoya City University Graduate School of Pharmaceutical Sciences, Nagoya, Japan. FAU - Sekiya, Takeo AU - Sekiya T AD - Department of Orthodontics, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan. AD - Department of Molecular Neurobiology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan. FAU - Kakita, Hiroki AU - Kakita H AD - Department of Molecular Neurobiology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan. AD - Department of Perinatal and Neonatal Medicine, Aichi Medical University, Nagakute, Japan. FAU - Waguri-Nagaya, Yuko AU - Waguri-Nagaya Y AD - Department of Joint Surgery for Rheumatic Diseases, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan. FAU - Miyazawa, Ken AU - Miyazawa K AD - Department of Orthodontics, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan. FAU - Asai, Kiyofumi AU - Asai K AD - Department of Molecular Neurobiology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan. FAU - Goto, Shigemi AU - Goto S AD - Department of Orthodontics, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20160708 PL - England TA - Stem Cells JT - Stem cells (Dayton, Ohio) JID - 9304532 RN - 0 (Antibodies) RN - 0 (CXCR4 protein, mouse) RN - 0 (Chemokine CX3CL1) RN - 0 (Chemokine CXCL12) RN - 0 (Chemokines, CXC) RN - 0 (Cx3cl1 protein, mouse) RN - 0 (Cxcl12 protein, mouse) RN - 0 (Cxcl7 protein, mouse) RN - 0 (RANK Ligand) RN - 0 (Receptor Activator of Nuclear Factor-kappa B) RN - 0 (Receptors, CXCR4) RN - 0 (Tnfrsf11a protein, mouse) RN - 0 (Tnfsf11 protein, mouse) RN - 81627-83-0 (Macrophage Colony-Stimulating Factor) RN - EC 3.1.3.48 (Leukocyte Common Antigens) RN - EC 3.1.3.48 (Ptprc protein, mouse) SB - IM MH - Animals MH - Antibodies/pharmacology MH - Bone Marrow Cells/cytology/drug effects/metabolism MH - Cell Differentiation MH - Chemokine CX3CL1/*genetics/metabolism MH - Chemokine CXCL12/antagonists & inhibitors/*genetics/metabolism MH - Chemokines, CXC/*genetics/metabolism MH - Flow Cytometry MH - Gene Expression Regulation MH - Leukocyte Common Antigens/deficiency/*genetics MH - Macrophage Colony-Stimulating Factor/pharmacology MH - Male MH - Mice MH - Osteoblasts/cytology/drug effects/*metabolism MH - Osteoclasts/cytology/drug effects/*metabolism MH - Osteogenesis/genetics MH - Primary Cell Culture MH - RANK Ligand/genetics/metabolism/pharmacology MH - Receptor Activator of Nuclear Factor-kappa B/genetics/metabolism MH - Receptors, CXCR4/*genetics/metabolism MH - Signal Transduction MH - Tibia/cytology/drug effects/metabolism OTO - NOTNLM OT - CXCR4+CD45- cells OT - Microenvironment OT - Osteoclastogenesis OT - SDF-1 OT - SDF-1/CXCR4 axis EDAT- 2016/06/25 06:00 MHDA- 2018/01/26 06:00 CRDT- 2016/06/25 06:00 PHST- 2015/11/30 00:00 [received] PHST- 2016/05/19 00:00 [revised] PHST- 2016/05/30 00:00 [accepted] PHST- 2016/06/25 06:00 [pubmed] PHST- 2018/01/26 06:00 [medline] PHST- 2016/06/25 06:00 [entrez] AID - 10.1002/stem.2440 [doi] PST - ppublish SO - Stem Cells. 2016 Nov;34(11):2733-2743. doi: 10.1002/stem.2440. Epub 2016 Jul 8.