{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T11:59:13Z","timestamp":1781265553442,"version":"3.54.1"},"posted":{"date-parts":[[2026,6,9]]},"group-title":"Immunology","reference-count":48,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T00:00:00Z","timestamp":1780963200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"name":"National Cancer Institute","award":["ZIA BC 011975"],"award-info":[{"award-number":["ZIA BC 011975"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,6,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Conventional type 1 dendritic cells (cDC1) specialize in cross-presentation and interleukin-12 production and are critical for immunity against intracellular pathogens and tumors, but remain rare\n                  <jats:italic>in vivo<\/jats:italic>\n                  , limiting mechanistic and translational studies. Existing bone marrow-derived dendritic cell (BMDC) methods do not achieve highly selective enrichment of cDC1 or scalable production at high purity. Here, we established a novel\n                  <jats:italic>in vitro<\/jats:italic>\n                  culture system for selective generation of CD103+ cDC1 from mouse bone marrow using defined media conditions together with recombinant FLT3L, GM-CSF, and Kit ligand (KitL), termed iDC1. iDC1 cultures enabled scalable generation of an estimated 1.5 x 10\n                  <jats:sup>9<\/jats:sup>\n                  CD103+ cDC1 at greater than 95% purity from a single mouse, representing at least a 75-fold increase relative to previous recombinant cytokine-based methods. Phenotypic and transcriptional analyses demonstrated that iDC1 closely align with the CD103+ cDC1 lineage while remaining clearly distinct from macrophage populations. Functionally, iDC1 responded robustly to innate stimulation, produced interleukin-12 and inflammatory chemokines, and efficiently cross-presented cell-associated antigen to CD8+ T cells. Mechanistically, KitL and GM-CSF regulated distinct stages of cDC1 generation, whereas proteomic, phospho-proteomic, and functional analyses demonstrated that GM-CSF suppresses apoptosis and oxidative stress while promoting cDC1 proliferation. iDC1 generation was dependent on the +32 kb\n                  <jats:italic>Irf8<\/jats:italic>\n                  enhancer required for\n                  <jats:italic>bona fide<\/jats:italic>\n                  cDC1 development, and STAT5-and BRD4-associated regulatory programs were identified as important regulators of efficient iDC1 generation. Together, these findings establish iDC1 cultures as a scalable platform for studying cDC1 biology and developing cDC1-based immunotherapeutic strategies.\n                <\/jats:p>","DOI":"10.64898\/2026.06.04.730212","type":"posted-content","created":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T19:10:18Z","timestamp":1781032218000},"source":"Crossref","is-referenced-by-count":0,"title":["Scalable generation of pure CD103+ cDC1 from iDC1 cultures"],"prefix":"10.64898","author":[{"given":"Sukriti","family":"Sharma","sequence":"first","affiliation":[{"name":"Experimental Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health; Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fiona","family":"Flynn","sequence":"additional","affiliation":[{"name":"Experimental Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health; Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian","family":"Capaldo","sequence":"additional","affiliation":[{"name":"Computational Genomics and Bioinformatics Branch, National Cancer Institute, National Institutes of Health, Frederick, MD 20850, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronald","family":"Holewinski","sequence":"additional","affiliation":[{"name":"Protein and Metabolite Characterization Core, Cancer Research and Technology Program, Frederick National Laboratory for Cancer Research, National Institutes of Health, Frederick, MD 21701, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingrong","family":"Chen","sequence":"additional","affiliation":[{"name":"Computational Genomics and Bioinformatics Branch, National Cancer Institute, National Institutes of Health, Frederick, MD 20850, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daoud","family":"Meerzaman","sequence":"additional","affiliation":[{"name":"Computational Genomics and Bioinformatics Branch, National Cancer Institute, National Institutes of Health, Frederick, MD 20850, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thorkell","family":"Andresson","sequence":"additional","affiliation":[{"name":"Protein and Metabolite Characterization Core, Cancer Research and Technology Program, Frederick National Laboratory for Cancer Research, National Institutes of Health, Frederick, MD 21701, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian T.","family":"Mayer","sequence":"additional","affiliation":[{"name":"Experimental Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health; Bethesda, MD 20892, 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