{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T13:07:28Z","timestamp":1790341648729,"version":"4.1.0"},"reference-count":48,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,3,19]],"date-time":"2024-03-19T00:00:00Z","timestamp":1710806400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,3,19]],"date-time":"2024-03-19T00:00:00Z","timestamp":1710806400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["424252458"],"award-info":[{"award-number":["424252458"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Walter Schulz Stiftung"},{"name":"Universit\u00e4tsklinikum M\u00fcnster"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Biol Proced Online"],"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n                <jats:title>Background<\/jats:title>\n                <jats:p>Osteoclasts are the tissue-specific macrophage population of the bone and unique in their bone-resorbing activity. Hence, they are fundamental for bone physiology in health and disease. However, efficient protocols for the isolation and study of primary human osteoclasts are scarce. In this study, we aimed to establish a protocol, which enables the efficient differentiation of functional human osteoclasts from monocytes.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Human monocytes were isolated through a double-density gradient from donor blood. Compared to standard differentiation schemes in polystyrene cell culture dishes, the yield of multinuclear osteoclasts was significantly increased upon initial differentiation of monocytes to macrophages in fluorinated ethylene propylene (FEP) Teflon bags. This initial differentiation phase was then followed by the development of terminal osteoclasts by addition of Receptor Activator of NF-\u03baB Ligand (RANKL). High concentrations of RANKL and Macrophage colony-stimulating factor (M-CSF) as well as an intermediate cell density further supported efficient cell differentiation. The generated cells were highly positive for CD45, CD14 as well as the osteoclast markers CD51\/<jats:italic>ITGAV<\/jats:italic> and Cathepsin K\/<jats:italic>CTSK<\/jats:italic>, thus identifying them as osteoclasts. The bone resorption of the osteoclasts was significantly increased when the cells were differentiated from macrophages derived from Teflon bags compared to macrophages derived from conventional cell culture plates.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusion<\/jats:title>\n                <jats:p>Our study has established a novel protocol for the isolation of primary human osteoclasts that improves osteoclastogenesis in comparison to the conventionally used cultivation approach.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s12575-024-00233-6","type":"journal-article","created":{"date-parts":[[2024,3,19]],"date-time":"2024-03-19T08:02:21Z","timestamp":1710835341000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["A novel method to efficiently differentiate human osteoclasts from blood-derived monocytes"],"prefix":"10.1186","volume":"26","author":[{"given":"Suganja","family":"Chandrabalan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linh","family":"Dang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Uwe","family":"Hansen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melanie","family":"Timmen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Corinna","family":"Wehmeyer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard","family":"Stange","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tim","family":"Bei\u00dfbarth","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Binder","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annalen","family":"Bleckmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kerstin","family":"Menck","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,3,19]]},"reference":[{"key":"233_CR1","doi-asserted-by":"publisher","first-page":"e10539","DOI":"10.1002\/jbm4.10539","volume":"5","author":"MM McDonald","year":"2021","unstructured":"McDonald MM, Kim AS, Mulholland BS, Rauner M. New insights into osteoclast biology. JBMR Plus. 2021;5:e10539.","journal-title":"JBMR Plus"},{"key":"233_CR2","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1002\/jcb.240550303","volume":"55","author":"AM Parfitt","year":"1994","unstructured":"Parfitt AM. Osteonal and hemi-osteonal remodeling: The spatial and temporal framework for signal traffic in adult human bone. J Cell Biochem. 1994;55:273\u201386.","journal-title":"J Cell Biochem"},{"key":"233_CR3","doi-asserted-by":"publisher","first-page":"1504","DOI":"10.1126\/science.289.5484.1504","volume":"289","author":"SL Teitelbaum","year":"2000","unstructured":"Teitelbaum SL. Bone resorption by osteoclasts. Science. 2000;289:1504\u20138.","journal-title":"Science"},{"key":"233_CR4","doi-asserted-by":"publisher","first-page":"337","DOI":"10.1038\/nature01658","volume":"423","author":"WJ Boyle","year":"2003","unstructured":"Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature. 2003;423:337\u201342.","journal-title":"Nature"},{"issue":"Suppl 2","key":"233_CR5","doi-asserted-by":"publisher","first-page":"ii32","DOI":"10.1136\/ard.61.suppl_2.ii32","volume":"61","author":"DH Jones","year":"2002","unstructured":"Jones DH, Kong YY, Penninger JM. Role of RANKL and RANK in bone loss and arthritis. Ann Rheum Dis. 2002;61(Suppl 2):ii32-39.","journal-title":"Ann Rheum Dis"},{"key":"233_CR6","doi-asserted-by":"publisher","first-page":"1507","DOI":"10.1002\/jbmr.5650081214","volume":"8","author":"EC Weir","year":"1993","unstructured":"Weir EC, Horowitz MC, Baron R, Centrella M, Kacinski BM, Insogna KL. Macrophage colony-stimulating factor release and receptor expression in bone cells. J Bone Miner Res. 1993;8:1507\u201318.","journal-title":"J Bone Miner Res"},{"key":"233_CR7","doi-asserted-by":"publisher","first-page":"1733","DOI":"10.1084\/jem.178.5.1733","volume":"178","author":"K Fuller","year":"1993","unstructured":"Fuller K, Owens JM, Jagger CJ, Wilson A, Moss R, Chambers TJ. Macrophage colony-stimulating factor stimulates survival and chemotactic behavior in isolated osteoclasts. J Exp Med. 1993;178:1733\u201344.","journal-title":"J Exp Med"},{"key":"233_CR8","doi-asserted-by":"publisher","first-page":"543","DOI":"10.1078\/0171-2985-00091","volume":"204","author":"J Xaus","year":"2001","unstructured":"Xaus J, Comalada M, Valledor AF, Card\u00f3 M, Herrero C, Soler C, Lloberas J, Celada A. Molecular mechanisms involved in macrophage survival, proliferation, activation or apoptosis. Immunobiology. 2001;204:543\u201350.","journal-title":"Immunobiology"},{"key":"233_CR9","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1007\/978-1-61779-415-5_13","volume":"816","author":"P Collin-Osdoby","year":"2012","unstructured":"Collin-Osdoby P, Osdoby P. RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol Biol. 2012;816:187\u2013202.","journal-title":"Methods Mol Biol"},{"key":"233_CR10","doi-asserted-by":"publisher","first-page":"1373","DOI":"10.1210\/endo-122-4-1373","volume":"122","author":"N Takahashi","year":"1988","unstructured":"Takahashi N, Yamana H, Yoshiki S, Roodman GD, Mundy GR, Jones SJ, Boyde A, Suda T. Osteoclast-like cell formation and its regulation by osteotropic hormones in mouse bone marrow cultures. Endocrinology. 1988;122:1373\u201382.","journal-title":"Endocrinology"},{"key":"233_CR11","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1038\/cmi.2013.19","volume":"10","author":"AM Burkhardt","year":"2013","unstructured":"Burkhardt AM, Zlotnik A. Translating translational research: mouse models of human disease. Cell Mol Immunol. 2013;10:373\u20134.","journal-title":"Cell Mol Immunol"},{"key":"233_CR12","doi-asserted-by":"publisher","first-page":"8380","DOI":"10.3892\/mmr.2017.7625","volume":"16","author":"ZH Li","year":"2017","unstructured":"Li ZH, Si Y, Xu G, Chen XM, Xiong H, Lai L, Zheng YQ, Zhang ZG. High-dose PMA with RANKL and MCSF induces THP-1 cell differentiation into human functional osteoclasts in vitro. Mol Med Rep. 2017;16:8380\u20134.","journal-title":"Mol Med Rep"},{"key":"233_CR13","doi-asserted-by":"publisher","first-page":"101651","DOI":"10.1016\/j.bonr.2022.101651","volume":"18","author":"SJA Remmers","year":"2023","unstructured":"Remmers SJA, van der Heijden FC, Ito K, Hofmann S. The effects of seeding density and osteoclastic supplement concentration on osteoclastic differentiation and resorption. Bone Rep. 2023;18:101651.","journal-title":"Bone Rep"},{"key":"233_CR14","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1007\/978-1-61779-415-5_11","volume":"816","author":"K Henriksen","year":"2012","unstructured":"Henriksen K, Karsdal MA, Taylor A, Tosh D, Coxon FP. Generation of human osteoclasts from peripheral blood. Methods Mol Biol. 2012;816:159\u201375.","journal-title":"Methods Mol Biol"},{"key":"233_CR15","doi-asserted-by":"crossref","unstructured":"Menck K, Behme D, Pantke M, Reiling N, Binder C, Pukrop T, Klemm F. Isolation of human monocytes by double gradient centrifugation and their differentiation to macrophages in teflon-coated cell culture bags. J Vis Exp. 2014;91:e51554.","DOI":"10.3791\/51554"},{"key":"233_CR16","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1186\/2049-2618-2-15","volume":"2","author":"AD Fernandes","year":"2014","unstructured":"Fernandes AD, Reid JN, Macklaim JM, McMurrough TA, Edgell DR, Gloor GB. Unifying the analysis of high-throughput sequencing datasets: characterizing RNA-seq, 16S rRNA gene sequencing and selective growth experiments by compositional data analysis. Microbiome. 2014;2:15.","journal-title":"Microbiome"},{"key":"233_CR17","doi-asserted-by":"publisher","first-page":"e74","DOI":"10.1182\/blood-2010-02-258558","volume":"116","author":"L Ziegler-Heitbrock","year":"2010","unstructured":"Ziegler-Heitbrock L, Ancuta P, Crowe S, Dalod M, Grau V, Hart DN, Leenen PJM, Liu Y-J, MacPherson G, Randolph GJ, et al. Nomenclature of monocytes and dendritic cells in blood. Blood. 2010;116:e74-80.","journal-title":"Blood"},{"key":"233_CR18","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/boneres.2015.32","volume":"3","author":"DM Merrild","year":"2015","unstructured":"Merrild DM, Pirapaharan DC, Andreasen CM, Kj\u00e6rsgaard-Andersen P, M\u00f8ller AM, Ding M, Delaiss\u00e9 JM, S\u00f8e K. Pit- and trench-forming osteoclasts: a distinction that matters. Bone Res. 2015;3:1\u201311.","journal-title":"Bone Res"},{"key":"233_CR19","doi-asserted-by":"publisher","first-page":"1477","DOI":"10.1359\/jbmr.2000.15.8.1477","volume":"15","author":"S Takeshita","year":"2000","unstructured":"Takeshita S, Kaji K, Kudo A. Identification and characterization of the new osteoclast progenitor with macrophage phenotypes being able to differentiate into mature osteoclasts. J Bone Miner Res. 2000;15:1477\u201388.","journal-title":"J Bone Miner Res"},{"key":"233_CR20","doi-asserted-by":"publisher","first-page":"337","DOI":"10.1007\/s00774-007-0773-9","volume":"25","author":"I Nakamura","year":"2007","unstructured":"Nakamura I, Duong LT, Rodan SB, Rodan GA. Involvement of alpha(v)beta3 integrins in osteoclast function. J Bone Miner Metab. 2007;25:337\u201344.","journal-title":"J Bone Miner Metab"},{"key":"233_CR21","doi-asserted-by":"publisher","first-page":"2402","DOI":"10.1002\/jcp.25389","volume":"231","author":"YH Chiu","year":"2016","unstructured":"Chiu YH, Ritchlin C. DC-STAMP: a key regulator in osteoclast differentiation. J Cell Physiol. 2016;231:2402\u20137.","journal-title":"J Cell Physiol"},{"key":"233_CR22","doi-asserted-by":"publisher","first-page":"233","DOI":"10.11005\/jbm.2014.21.4.233","volume":"21","author":"JH Kim","year":"2014","unstructured":"Kim JH, Kim N. Regulation of NFATc1 in osteoclast differentiation. J Bone Metab. 2014;21:233\u201341.","journal-title":"J Bone Metab"},{"key":"233_CR23","doi-asserted-by":"publisher","first-page":"205","DOI":"10.1016\/S0928-4931(98)00052-6","volume":"6","author":"HK V\u00e4\u00e4n\u00e4nen","year":"1998","unstructured":"V\u00e4\u00e4n\u00e4nen HK, Liu Y, Lehenkari P, Uemara T. How do osteoclasts resorb bone? Mater Sci Eng, C. 1998;6:205\u20139.","journal-title":"Mater Sci Eng, C"},{"key":"233_CR24","doi-asserted-by":"publisher","first-page":"eaaw6143","DOI":"10.1126\/scitranslmed.aaw6143","volume":"12","author":"L Zhu","year":"2020","unstructured":"Zhu L, Tang Y, Li XY, Keller ET, Yang J, Cho J-S, Feinberg TY, Weiss SJ. Osteoclast-mediated bone resorption is controlled by a compensatory network of secreted and membrane-tethered metalloproteinases. Sci Transl Med. 2020;12:eaaw6143.","journal-title":"Sci Transl Med"},{"key":"233_CR25","doi-asserted-by":"publisher","first-page":"128","DOI":"10.1006\/excr.1998.4071","volume":"242","author":"P Lehenkari","year":"1998","unstructured":"Lehenkari P, Hentunen TA, Laitala-Leinonen T, Tuukkanen J, V\u00e4\u00e4n\u00e4nen HK. Carbonic anhydrase II plays a major role in osteoclast differentiation and bone resorption by effecting the steady state Intracellular pH and Ca2+. Exp Cell Res. 1998;242:128\u201337.","journal-title":"Exp Cell Res"},{"key":"233_CR26","doi-asserted-by":"publisher","first-page":"e0277871","DOI":"10.1371\/journal.pone.0277871","volume":"17","author":"Y Cheng","year":"2022","unstructured":"Cheng Y, Liu H, Li J, Ma Y, Song C, Wang Y, Li P, Chen Y, Zhang Z. Evaluation of culture conditions for osteoclastogenesis in RAW264.7 cells. PLOS ONE. 2022;17:e0277871.","journal-title":"PLOS ONE"},{"key":"233_CR27","doi-asserted-by":"crossref","unstructured":"Wang L, Zheng S, Guo Y, Pan Y, Sun J, Xu W, Lu J, Li W, Ma Y. Isolation, purification, and differentiation of osteoclast precursors from rat bone marrow. J Vis Exp.\u00a02019;147:e58895.","DOI":"10.3791\/58895-v"},{"key":"233_CR28","doi-asserted-by":"publisher","first-page":"6","DOI":"10.3390\/jdb10010006","volume":"10","author":"DR Halloran","year":"2022","unstructured":"Halloran DR, Heubel B, MacMurray C, Root D, Eskander M, McTague SP, Pelkey H, Nohe A. Differentiation of cells isolated from human femoral heads into functional osteoclasts. Journal of Developmental Biology. 2022;10:6.","journal-title":"Journal of Developmental Biology"},{"key":"233_CR29","doi-asserted-by":"publisher","first-page":"2567","DOI":"10.3389\/fimmu.2018.02567","volume":"9","author":"MB Madel","year":"2018","unstructured":"Madel MB, Ib\u00e1\u00f1ez L, Rouleau M, Wakkach A, Blin-Wakkach C. A novel reliable and efficient procedure for purification of mature osteoclasts allowing functional assays in mouse cells. Front Immunol. 2018;9:2567.","journal-title":"Front Immunol"},{"key":"233_CR30","doi-asserted-by":"publisher","first-page":"6","DOI":"10.1186\/1479-5876-2-6","volume":"2","author":"M Susa","year":"2004","unstructured":"Susa M, Luong-Nguyen NH, Cappellen D, Zamurovic N, Gamse R. Human primary osteoclasts: in vitro generation and applications as pharmacological and clinical assay. J Transl Med. 2004;2:6.","journal-title":"J Transl Med"},{"key":"233_CR31","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1042\/cs0990133","volume":"99","author":"GC Nicholson","year":"2000","unstructured":"Nicholson GC, Malakellis M, Collier FM, Cameron PU, Holloway WR, Gough TJ, Gregorio-King C, Kirkland MA, Myers DE. Induction of osteoclasts from CD14-positive human peripheral blood mononuclear cells by receptor activator of nuclear factor kappaB ligand (RANKL). Clin Sci (Lond). 2000;99:133\u201340.","journal-title":"Clin Sci (Lond)"},{"key":"233_CR32","doi-asserted-by":"publisher","first-page":"2045","DOI":"10.12688\/f1000research.12802.3","volume":"6","author":"J Bhattacharjee","year":"2018","unstructured":"Bhattacharjee J, Das B, Mishra A, Sahay P, Upadhyay P. Monocytes isolated by positive and negative magnetic sorting techniques show different molecular characteristics and immunophenotypic behaviour. F1000Res. 2018;6:2045.","journal-title":"F1000Res"},{"key":"233_CR33","doi-asserted-by":"publisher","first-page":"1583","DOI":"10.3390\/biology11111583","volume":"11","author":"M Hornschuh","year":"2022","unstructured":"Hornschuh M, Haas V, Winkel PP, G\u00f6kyildirim MY, Mullins CS, Wrobel IM, Manteuffel C, Wirthgen E. Negative magnetic sorting preserves the functionality of ex vivo cultivated non-adherent human monocytes. Biology. 2022;11:1583.","journal-title":"Biology"},{"key":"233_CR34","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1111\/imm.13125","volume":"159","author":"MC Nielsen","year":"2020","unstructured":"Nielsen MC, Andersen MN, M\u00f8ller HJ. Monocyte isolation techniques significantly impact the phenotype of both isolated monocytes and derived macrophages in vitro. Immunology. 2020;159:63\u201374.","journal-title":"Immunology"},{"key":"233_CR35","doi-asserted-by":"publisher","first-page":"2773","DOI":"10.1111\/jcmm.13192","volume":"21","author":"M Becerikli","year":"2017","unstructured":"Becerikli M, Jaurich H, Schira J, Schulte M, D\u00f6bele C, Wallner C, Abraham S, Wagner JM, Dadras M, Kneser U, et al. Age-dependent alterations in osteoblast and osteoclast activity in human cancellous bone. J Cell Mol Med. 2017;21:2773\u201381.","journal-title":"J Cell Mol Med"},{"key":"233_CR36","doi-asserted-by":"publisher","first-page":"673","DOI":"10.1677\/joe.0.1720673","volume":"172","author":"M Jevon","year":"2002","unstructured":"Jevon M, Sabokbar A, Fujikawa Y, Hirayama T, Neale SD, Wass J, Athanasou NA. Gender- and age-related differences in osteoclast formation from circulating precursors. J Endocrinol. 2002;172:673\u201381.","journal-title":"J Endocrinol"},{"key":"233_CR37","doi-asserted-by":"publisher","first-page":"1800","DOI":"10.1111\/trf.14621","volume":"58","author":"N Fekete","year":"2018","unstructured":"Fekete N, B\u00e9land AV, Campbell K, Clark SL, Hoesli CA. Bags versus flasks: a comparison of cell culture systems for the production of dendritic cell\u2013based immunotherapies. Transfusion. 2018;58:1800\u201313.","journal-title":"Transfusion"},{"key":"233_CR38","first-page":"5","volume":"1","author":"HJ Busscher","year":"1991","unstructured":"Busscher HJ, Stokroos I, Golverdingen JG, Shakenraad JM. Adhesion and spreading of human fibroblasts on superhydrophobic Fep-Teflon. Cells and Materials. 1991;1:5.","journal-title":"Cells and Materials"},{"key":"233_CR39","first-page":"617","volume":"47","author":"JW van der Meer","year":"1982","unstructured":"van der Meer JW, van de Gevel JS, van Oud B, Alblas A, Kramps JA, van Zwet TL, Leijh PC, van Furth R. Characteristics of human monocytes cultured in the Teflon culture bag. Immunology. 1982;47:617\u201325.","journal-title":"Immunology"},{"key":"233_CR40","doi-asserted-by":"publisher","first-page":"S195","DOI":"10.1016\/j.jcyt.2020.04.059","volume":"22","author":"L Zhang","year":"2020","unstructured":"Zhang L, Fekete N, Campbell K. 3D MSC Culture in Saint-Gobain VueLife\u00ae FEP Bags Using Microcarriers. Cytotherapy. 2020;22:S195\u20136.","journal-title":"Cytotherapy"},{"key":"233_CR41","doi-asserted-by":"publisher","first-page":"1431","DOI":"10.3727\/096368910X557164","volume":"20","author":"M Bouchentouf","year":"2011","unstructured":"Bouchentouf M, Forner K, Cuerquis J, Boulassel MR, Routy JP, Waller EK, Quyyumi AA, Paradis P, Schiffrin EL, Galipeau J. A novel and simplified method of culture of human blood-derived early endothelial progenitor cells for the treatment of ischemic vascular disease. Cell Transplant. 2011;20:1431\u201343.","journal-title":"Cell Transplant"},{"key":"233_CR42","doi-asserted-by":"publisher","first-page":"199","DOI":"10.1080\/10245330410001701486","volume":"9","author":"T B\u00fcchler","year":"2004","unstructured":"B\u00fcchler T, Kov\u00e1\u0159ov\u00e1 L, Musilov\u00e1 R, Bourkov\u00e1 L, O\u010dadl\u00edkov\u00e1 D, Bulikov\u00e1 A, Han\u00e1k L, Mich\u00e1lek J, H\u00e1jek R. Generation of dendritic cells using cell culture bags-description of a method and review of literature. Hematology. 2004;9:199\u2013205.","journal-title":"Hematology"},{"key":"233_CR43","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1016\/S0022-1759(02)00015-7","volume":"262","author":"CA Guyre","year":"2002","unstructured":"Guyre CA, Fisher JL, Waugh MG, Wallace PK, Tretter CG, Ernstoff MS, Barth RJ. Advantages of hydrophobic culture bags over flasks for the generation of monocyte-derived dendritic cells for clinical applications. J Immunol Methods. 2002;262:85\u201394.","journal-title":"J Immunol Methods"},{"key":"233_CR44","doi-asserted-by":"publisher","first-page":"374","DOI":"10.1097\/00002371-200307000-00010","volume":"26","author":"RV Sorg","year":"2003","unstructured":"Sorg RV, Ozcan Z, Brefort T, Fischer J, Ackermann R, M\u00fcller M, Wernet P. Clinical-scale generation of dendritic cells in a closed system. J Immunother. 2003;26:374\u201383.","journal-title":"J Immunother"},{"key":"233_CR45","doi-asserted-by":"publisher","first-page":"176","DOI":"10.1210\/me.2007-0237","volume":"22","author":"K Kim","year":"2008","unstructured":"Kim K, Lee SH, Ha Kim J, Choi Y, Kim N. NFATc1 induces osteoclast fusion via up-regulation of Atp6v0d2 and the dendritic cell-specific transmembrane protein (DC-STAMP). Mol Endocrinol. 2008;22:176\u201385.","journal-title":"Mol Endocrinol"},{"key":"233_CR46","doi-asserted-by":"publisher","first-page":"2538","DOI":"10.1002\/jcp.25638","volume":"232","author":"YH Chiu","year":"2017","unstructured":"Chiu YH, Schwarz E, Li D, Xu Y, Sheu TR, Li J, de Mesy Bentley KL, Feng C, Wang B, Wang JC, et al. Dendritic Cell-Specific Transmembrane Protein (DC-STAMP) regulates osteoclast differentiation via the Ca2+ \/NFATc1 Axis. J Cell Physiol. 2017;232:2538\u201349.","journal-title":"J Cell Physiol"},{"key":"233_CR47","doi-asserted-by":"publisher","first-page":"5379","DOI":"10.7150\/thno.28391","volume":"8","author":"X Duan","year":"2018","unstructured":"Duan X, Yang S, Zhang L, Yang T. V-ATPases and osteoclasts: ambiguous future of V-ATPases inhibitors in osteoporosis. Theranostics. 2018;8:5379\u201399.","journal-title":"Theranostics"},{"key":"233_CR48","doi-asserted-by":"crossref","unstructured":"Chou J, Chan MF, Werb Z. Metalloproteinases: a functional pathway for myeloid cells. 2nd ed. Microbiol Spectr.\u00a02016;4:10.1128.","DOI":"10.1128\/microbiolspec.MCHD-0002-2015"}],"container-title":["Biological Procedures Online"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12575-024-00233-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12575-024-00233-6\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12575-024-00233-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,3,19]],"date-time":"2024-03-19T08:02:46Z","timestamp":1710835366000},"score":1,"resource":{"primary":{"URL":"https:\/\/biologicalproceduresonline.biomedcentral.com\/articles\/10.1186\/s12575-024-00233-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,19]]},"references-count":48,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["233"],"URL":"https:\/\/doi.org\/10.1186\/s12575-024-00233-6","relation":{},"ISSN":["1480-9222"],"issn-type":[{"value":"1480-9222","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,3,19]]},"assertion":[{"value":"31 August 2023","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 March 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 March 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"This study has been approved by the local ethics committees of the Medical association Westfalen-Lippe (approval no. 2023\u2013475-f-N).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"7"}}