{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T18:18:58Z","timestamp":1787768338720,"version":"build-2784847793"},"reference-count":55,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T00:00:00Z","timestamp":1787702400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    The selection of an appropriate differentiation protocol for murine bone marrow-derived dendritic cells (BM-DCs) still presents a challenge due to the inherent plasticity of dendritic cells (DCs) and the high diversity of generation methods. As such, it is essential to evaluate\n                    <jats:italic>in vitro<\/jats:italic>\n                    BM-DCs characteristics during initial steps of experimental model development and tailor the protocols according to specific research goals while preserving the functional and phenotypic properties of DCs. In our study, we observed relevant variations in phenotypic and functional characteristics of BM-DCs differentiated with GM-CSF alone (GDCs) compared to those differentiated with GM-CSF and IL-4 (G4DCs). We found that GDCs exhibited high levels of costimulatory molecules, indicating a phenotypically mature state, but preserved the ability to respond to LPS (lipopolysaccharide) maturation stimuli by cytokine production. Moreover, cytokine secretion patterns observed in prime\/ boost co-culture systems indicate the potential of Ag-pulsed BM-DCs to prime naive T cells. In contrast, while addition of IL-4 resulted in cells with a more immature phenotype that are responsive to LPS-driven maturation, it also promoted nonspecific T cell signaling that could interfere with\n                    <jats:italic>in vitro<\/jats:italic>\n                    evaluation of priming and antigen-specific T cell responses. These results highlight the impact of specific differentiation conditions that impose on the phenotypic and functional patterns of BM-DCs. Furthermore, the study emphasizes the significance of reassessing, even widely, the already employed protocols for DCs differentiation, and moreover, our study can provide valuable insights and guidance for novel\n                    <jats:italic>in vitro<\/jats:italic>\n                    experimental designs based on BM-DCs.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0356806","type":"journal-article","created":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T17:30:57Z","timestamp":1787765457000},"page":"e0356806","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Phenotypic and functional patterns of murine bone marrow-derived dendritic cells influenced by in vitro differentiation conditions"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8848-7252","authenticated-orcid":true,"given":"Irina Elena","family":"Ionescu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catalin","family":"Tucureanu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raluca Elena","family":"Chelmus","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4052-2383","authenticated-orcid":true,"given":"Vlad Constantin","family":"Tofan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabiola Margareta","family":"Ionita","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrian","family":"Onu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica","family":"Neagu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Crina","family":"Stavaru","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iuliana","family":"Caras","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,8,26]]},"reference":[{"issue":"3","key":"pone.0356806.ref001","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1158\/2326-6066.CIR-13-0102","article-title":"Dendritic cells: master regulators of the immune response","volume":"1","author":"I Mellman","year":"2013","journal-title":"Cancer Immunol Res"},{"key":"pone.0356806.ref002","doi-asserted-by":"crossref","first-page":"924","DOI":"10.3389\/fimmu.2020.00924","article-title":"Dendritic cells and their role in immunotherapy","volume":"11","author":"A Gardner","year":"2020","journal-title":"Front Immunol"},{"issue":"7","key":"pone.0356806.ref003","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1038\/s41401-020-0415-5","article-title":"Ex vivo pulsed dendritic cell vaccination against cancer","volume":"41","author":"Y-Z Gu","year":"2020","journal-title":"Acta Pharmacol Sin"},{"key":"pone.0356806.ref004","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1146\/annurev-immunol-020711-074950","article-title":"The dendritic cell lineage: ontogeny and function of dendritic cells and their subsets in the steady state and the inflamed setting","volume":"31","author":"M Merad","year":"2013","journal-title":"Annu Rev Immunol"},{"issue":"2","key":"pone.0356806.ref005","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1038\/s41577-020-00413-x","article-title":"Genetic models of human and mouse dendritic cell development and function","volume":"21","author":"DA Anderson","year":"2021","journal-title":"Nat Rev Immunol"},{"key":"pone.0356806.ref006","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/j.copbio.2020.12.010","article-title":"Dendritic cell metabolism: moving beyond in vitro-culture-generated paradigms","volume":"68","author":"L Minarrieta","year":"2021","journal-title":"Curr Opin Biotechnol"},{"issue":"2","key":"pone.0356806.ref007","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/0022-1759(92)90199-4","article-title":"Dendritic cells from mouse bone marrow: in vitro differentiation using low doses of recombinant granulocyte-macrophage colony-stimulating factor","volume":"154","author":"C Scheicher","year":"1992","journal-title":"J Immunol Methods"},{"issue":"1","key":"pone.0356806.ref008","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1084\/jem.180.1.83","article-title":"Proliferating dendritic cell progenitors in human blood","volume":"180","author":"N Romani","year":"1994","journal-title":"J Exp Med"},{"key":"pone.0356806.ref009","first-page":"25","article-title":"In-vitro differentiation of mature dendritic cells from human blood monocytes","volume":"6","author":"R Gieseler","year":"1998","journal-title":"Dev Immunol"},{"issue":"12","key":"pone.0356806.ref010","doi-asserted-by":"crossref","first-page":"3658","DOI":"10.1016\/j.celrep.2018.05.068","article-title":"Notch signaling facilitates in vitro generation of cross-presenting classical dendritic cells","volume":"23","author":"ME Kirkling","year":"2018","journal-title":"Cell Rep"},{"issue":"10","key":"pone.0356806.ref011","doi-asserted-by":"crossref","first-page":"3006","DOI":"10.4049\/jimmunol.1701402","article-title":"Differentiation of langerhans cells from monocytes and their specific function in inducing IL-22-specific Th cells","volume":"201","author":"Y Otsuka","year":"2018","journal-title":"J Immunol"},{"issue":"7","key":"pone.0356806.ref012","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1038\/s41590-019-0420-3","article-title":"Plasmacytoid dendritic cells develop from Ly6D+ lymphoid progenitors distinct from the myeloid lineage","volume":"20","author":"RJ Dress","year":"2019","journal-title":"Nat Immunol"},{"issue":"12","key":"pone.0356806.ref013","doi-asserted-by":"crossref","first-page":"1946","DOI":"10.1002\/eji.202149663","article-title":"Flt3L, LIF, and IL-10 combination promotes the selective in vitro development of ESAMlow cDC2B from murine bone marrow","volume":"52","author":"L Cyran","year":"2022","journal-title":"Eur J Immunol"},{"issue":"2","key":"pone.0356806.ref014","doi-asserted-by":"crossref","first-page":"181","DOI":"10.3390\/cancers11020181","article-title":"Cord-blood-stem-cell-derived conventional dendritic cells specifically originate from CD115-expressing precursors","volume":"11","author":"M Plantinga","year":"2019","journal-title":"Cancers (Basel)"},{"key":"pone.0356806.ref015","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1016\/j.imbio.2007.09.001","article-title":"Factors influencing the generation of murine dendritic cells from bone marrow: the special role of fetal calf serum","volume":"212","author":"MB Lutz","year":"2007","journal-title":"Immunobiology"},{"issue":"1","key":"pone.0356806.ref016","doi-asserted-by":"crossref","first-page":"233","DOI":"10.3892\/mmr.2015.3349","article-title":"Interleukin-4 affects the mature phenotype and function of rat bone marrow-derived dendritic cells","volume":"12","author":"S Wang","year":"2015","journal-title":"Mol Med Rep"},{"key":"pone.0356806.ref017","doi-asserted-by":"crossref","first-page":"2352","DOI":"10.3389\/fimmu.2019.02352","article-title":"The culture dish surface influences the phenotype and cytokine production of human monocyte-derived dendritic cells","volume":"10","author":"A Sauter","year":"2019","journal-title":"Front Immunol"},{"issue":"2","key":"pone.0356806.ref018","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1039\/D1LC00876E","article-title":"Exhausted mature dendritic cells exhibit a slower and less persistent random motility but retain chemotaxis against CCL19","volume":"22","author":"Y Choi","year":"2022","journal-title":"Lab Chip"},{"issue":"4","key":"pone.0356806.ref019","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1038\/79758","article-title":"Kinetics of dendritic cell activation: impact on priming of TH1, TH2 and nonpolarized T cells","volume":"1","author":"A Langenkamp","year":"2000","journal-title":"Nat Immunol"},{"issue":"6","key":"pone.0356806.ref020","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1084\/jem.176.6.1693","article-title":"Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte\/macrophage colony-stimulating factor","volume":"176","author":"K Inaba","year":"1992","journal-title":"J Exp Med"},{"issue":"1","key":"pone.0356806.ref021","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/S0022-1759(98)00204-X","article-title":"An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow","volume":"223","author":"MB Lutz","year":"1999","journal-title":"J Immunol Methods"},{"issue":"4","key":"pone.0356806.ref022","doi-asserted-by":"crossref","first-page":"1109","DOI":"10.1084\/jem.179.4.1109","article-title":"Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte\/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha","volume":"179","author":"F Sallusto","year":"1994","journal-title":"J Exp Med"},{"issue":"7","key":"pone.0356806.ref023","doi-asserted-by":"crossref","first-page":"1813","DOI":"10.1002\/1521-4141(200007)30:7<1813::AID-IMMU1813>3.0.CO;2-8","article-title":"Immature dendritic cells generated with low doses of GM-CSF in the absence of IL-4 are maturation resistant and prolong allograft survival in vivo","volume":"30","author":"MB Lutz","year":"2000","journal-title":"Eur J Immunol"},{"key":"pone.0356806.ref024","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/S0022-1759(02)00013-3","article-title":"A novel bulk-culture method for generating mature dendritic cells from mouse bone marrow cells","volume":"262","author":"Y-I Son","year":"2002","journal-title":"J Immunol Methods"},{"issue":"4","key":"pone.0356806.ref025","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1189\/jlb.0804473","article-title":"IL-4 supports the generation of a dendritic cell subset from murine bone marrow with altered endocytosis capacity","volume":"77","author":"M Menges","year":"2005","journal-title":"J Leukoc Biol"},{"issue":"4","key":"pone.0356806.ref026","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1111\/j.1365-2567.2010.03324.x","article-title":"Innate immune stimuli modulate bone marrow-derived dendritic cell production in vitro by toll-like receptor-dependent and -independent mechanisms","volume":"131","author":"JE Downes","year":"2010","journal-title":"Immunology"},{"issue":"6","key":"pone.0356806.ref027","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1016\/j.immuni.2015.05.018","article-title":"GM-CSF mouse bone marrow cultures comprise a heterogeneous population of CD11c(+)MHCII(+) macrophages and dendritic cells","volume":"42","author":"J Helft","year":"2015","journal-title":"Immunity"},{"key":"pone.0356806.ref028","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.jconrel.2014.06.024","article-title":"Choose your models wisely: how different murine bone marrow-derived dendritic cell protocols influence the success of nanoparticulate vaccines in vitro","volume":"195","author":"H Dewitte","year":"2014","journal-title":"J Control Release"},{"issue":"6","key":"pone.0356806.ref029","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1084\/jem.192.6.823","article-title":"Interleukin (IL)-4 is a major regulatory cytokine governing bioactive IL-12 production by mouse and human dendritic cells","volume":"192","author":"H Hochrein","year":"2000","journal-title":"J Exp Med"},{"issue":"1","key":"pone.0356806.ref030","doi-asserted-by":"crossref","first-page":"168","DOI":"10.4049\/jimmunol.162.1.168","article-title":"Generation of tumor immunity by bone marrow-derived dendritic cells correlates with dendritic cell maturation stage","volume":"162","author":"MS Labeur","year":"1999","journal-title":"J Immunol"},{"issue":"8","key":"pone.0356806.ref031","doi-asserted-by":"crossref","first-page":"2869","DOI":"10.1182\/blood.V99.8.2869","article-title":"Increased dendritic cell number and function following continuous in vivo infusion of granulocyte macrophage-colony-stimulating factor and interleukin-4","volume":"99","author":"SK Basak","year":"2002","journal-title":"Blood"},{"issue":"3","key":"pone.0356806.ref032","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1111\/j.1365-3083.2005.01556.x","article-title":"Influence of interleukin\u20104 on the phenotype and function of bone marrow\u2010derived murine dendritic cells generated under serum\u2010free conditions","volume":"61","author":"JW Wells","year":"2005","journal-title":"Scand J Immunol"},{"issue":"20","key":"pone.0356806.ref033","article-title":"Immune cell isolation from mouse femur bone marrow","volume":"5","author":"X Liu","year":"2015","journal-title":"Bio Protoc"},{"issue":"10","key":"pone.0356806.ref034","doi-asserted-by":"crossref","first-page":"1520","DOI":"10.3390\/vaccines11101520","article-title":"Generation of a DSF-guided refolded bacterially expressed hemagglutinin ectodomain of influenza virus A\/Puerto Rico\/8\/1934 H1N1 as a model for influenza vaccine antigens","volume":"11","author":"V-C Tofan","year":"2023","journal-title":"Vaccines (Basel)"},{"issue":"1","key":"pone.0356806.ref035","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1146\/annurev-immunol-042617-053352","article-title":"Signaling and function of interleukin-2 in T lymphocytes","volume":"36","author":"SH Ross","year":"2018","journal-title":"Annu Rev Immunol"},{"key":"pone.0356806.ref036","volume-title":"R: a language and environment for statistical computing, version 4.6.1","author":"R Core Team","year":"2024"},{"key":"pone.0356806.ref037","volume-title":"RStudio: integrated development environment for R, version 2023.09.1+494","author":"Posit Team","year":"2023"},{"key":"pone.0356806.ref038","doi-asserted-by":"crossref","unstructured":"Wickham H, Chang W, Henry L, et al. ggplot2: Create elegant data visualisations using the grammar of graphics, 4.0.2; 2007. doi: 10.32614\/CRAN.package.ggplot2","DOI":"10.32614\/CRAN.package.ggplot2"},{"issue":"3","key":"pone.0356806.ref039","article-title":"Complex heatmap visualization","volume":"1","author":"Z Gu","year":"2022","journal-title":"iMeta"},{"issue":"22","key":"pone.0356806.ref040","doi-asserted-by":"crossref","first-page":"12011","DOI":"10.1128\/JVI.01316-10","article-title":"Dendritic cell activation by recombinant hemagglutinin proteins of H1N1 and H5N1 influenza A viruses","volume":"84","author":"W-C Liu","year":"2010","journal-title":"J Virol"},{"issue":"3","key":"pone.0356806.ref041","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1093\/intimm\/dxh384","article-title":"Purification of splenic dendritic cells induces maturation and capacity to stimulate Th1 response in vivo","volume":"18","author":"G Schlecht","year":"2006","journal-title":"Int Immunol"},{"issue":"4","key":"pone.0356806.ref042","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1016\/j.immuni.2007.08.015","article-title":"Disruption of E-cadherin-mediated adhesion induces a functionally distinct pathway of dendritic cell maturation","volume":"27","author":"A Jiang","year":"2007","journal-title":"Immunity"},{"issue":"10","key":"pone.0356806.ref043","doi-asserted-by":"crossref","first-page":"3129","DOI":"10.4049\/jimmunol.1800031","article-title":"GM-CSF culture revisited: preparation of bulk populations of highly pure dendritic cells from mouse bone marrow","volume":"201","author":"D Jin","year":"2018","journal-title":"J Immunol"},{"key":"pone.0356806.ref044","article-title":"Interleukin-4 inhibits regulatory T cell differentiation through regulating CD103 dendritic cells","volume":"8","author":"L Tu","year":"2017","journal-title":"Front Immunol"},{"issue":"7","key":"pone.0356806.ref045","doi-asserted-by":"crossref","first-page":"4312","DOI":"10.4049\/jimmunol.166.7.4312","article-title":"Regulatory activity of autocrine IL-10 on dendritic cell functions","volume":"166","author":"S Corinti","year":"2001","journal-title":"J Immunol"},{"issue":"5","key":"pone.0356806.ref046","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1016\/j.imbio.2006.02.001","article-title":"Identification of a rat bone marrow-derived dendritic cell population which secretes both IL-10 and IL-12: evidence against a reciprocal relationship between IL-10 and IL-12 secretion","volume":"211","author":"M Muthana","year":"2006","journal-title":"Immunobiology"},{"issue":"1","key":"pone.0356806.ref047","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.cyto.2015.07.026","article-title":"Making many from few: IL-12p40 as a model for the combinatorial assembly of heterodimeric cytokines","volume":"76","author":"K Abdi","year":"2015","journal-title":"Cytokine"},{"issue":"7","key":"pone.0356806.ref048","doi-asserted-by":"crossref","first-page":"3645","DOI":"10.4049\/jimmunol.171.7.3645","article-title":"Lipopolysaccharide-induced suppression of airway Th2 responses does not require IL-12 production by dendritic cells","volume":"171","author":"H Kuipers","year":"2003","journal-title":"J Immunol"},{"issue":"9","key":"pone.0356806.ref049","doi-asserted-by":"crossref","first-page":"1323","DOI":"10.4161\/hv.21350","article-title":"Dendritic cell-based in vitro assays for vaccine immunogenicity","volume":"8","author":"R Vandebriel","year":"2012","journal-title":"Hum Vaccin Immunother"},{"issue":"8","key":"pone.0356806.ref050","doi-asserted-by":"crossref","first-page":"929","DOI":"10.3390\/vaccines9080929","article-title":"In vitro priming of human T cells by dendritic cells provides a screening tool for candidate vaccines for Burkholderia pseudomallei","volume":"9","author":"D Reddi","year":"2021","journal-title":"Vaccines (Basel)"},{"key":"pone.0356806.ref051","doi-asserted-by":"crossref","first-page":"1419","DOI":"10.3389\/fimmu.2017.01419","article-title":"Human CD141+ dendritic cell and CD1c+ dendritic cell undergo concordant early genetic programming after activation in humanized mice in vivo","volume":"8","author":"Y Minoda","year":"2017","journal-title":"Front Immunol"},{"issue":"1","key":"pone.0356806.ref052","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1084\/jem.20072665","article-title":"Alarming dendritic cells for Th2 induction","volume":"205","author":"AS MacDonald","year":"2008","journal-title":"J Exp Med"},{"issue":"2","key":"pone.0356806.ref053","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1111\/j.1365-2567.2005.02305.x","article-title":"Interleukin-4 can induce interleukin-4 production in dendritic cells","volume":"117","author":"A Maroof","year":"2006","journal-title":"Immunology"},{"issue":"3","key":"pone.0356806.ref054","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1189\/jlb.1104631","article-title":"Polarization of naive T cells into Th1 or Th2 by distinct cytokine-driven murine dendritic cell populations: implications for immunotherapy","volume":"78","author":"M Feili-Hariri","year":"2005","journal-title":"J Leukoc Biol"},{"issue":"7","key":"pone.0356806.ref055","doi-asserted-by":"crossref","first-page":"2021","DOI":"10.1002\/1521-4141(200207)32:7<2021::AID-IMMU2021>3.0.CO;2-J","article-title":"Regulatory Th2 response induced following adoptive transfer of dendritic cells in prediabetic NOD mice","volume":"32","author":"M Feili-Hariri","year":"2002","journal-title":"Eur J Immunol"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0356806","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T17:31:08Z","timestamp":1787765468000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0356806"}},"subtitle":[],"editor":[{"given":"Francesco","family":"Bertolini","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,8,26]]},"references-count":55,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8,26]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0356806","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,26]]}}}