Scalable generation of pure CD103+ cDC1 from iDC1 cultures
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Scalable generation of pure CD103+ cDC1 from iDC1 cultures

Sukriti Sharma
1
,
Fiona Flynn
1
,
Brian Capaldo
2
,
Ronald Holewinski
3
,
Qingrong Chen
2
,
Daoud Meerzaman
2
,
Thorkell Andresson
3
,
Christian T. Mayer
1,*
*Correspondence to: Christian T. Mayer, Experimental Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. E-mail: christian.mayer@nih.gov
Myeloid Cells. 2026;1:202613. 10.70401/mc.2026.0012
Received: June 04, 2026Accepted: August 28, 2026Published: August 28, 2026
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This manuscript is made available in its unedited form to allow early access to the reported findings. Further editing will be completed before final publication. As such, the content may include errors, and standard legal disclaimers are applicable.

Abstract

Aims: 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 in vivo, limiting mechanistic and translational studies. Existing bone marrow-derived dendritic cell (BMDC) methods do not achieve selective enrichment of cDC1 or scalable production at high purity. We therefore aimed to establish a high-efficiency BMDC culture system for selective generation of CD103+ cDC1 from mouse bone marrow.

Methods: Mouse bone marrow cells were cultured in defined medium with recombinant FLT3L, low-dose GM-CSF, and Kit ligand (KitL) to generate induced cDC1 (iDC1). Phenotypic, transcriptomic, proteomic, phospho-proteomic, and functional analyses were performed and compared with established BMDC methods and ex vivo immune cell populations.

Results: iDC1 cultures enabled scalable generation of an estimated 1.5 × 109 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. iDC1 closely aligned with the cDC1 lineage while distinct from macrophages. Functionally, iDC1 responded robustly to innate stimulation, secreted IL-12p40 and inflammatory chemokines, and efficiently cross-presented cell-associated antigen to CD8+ T cells. Mechanistically, KitL and GM-CSF cooperated during early cDC1 generation, whereas GM-CSF promoted proliferation and survival during later stages of culture. iDC1 generation depended on the +32 kb Irf8 enhancer, and STAT5- and BRD4-associated regulatory programs contributed to efficient iDC1 generation.

Conclusion: iDC1 cultures are a scalable platform for studying cDC1 biology and may facilitate development and preclinical evaluation of cDC1-based immunotherapeutic strategies.

Keywords

iDC1, cDC1, CD103+ DC, GM-CSF, cross-presentation, STAT5, BRD4, IL-12p40

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Sharma S, Flynn F, Capaldo B, Holewinski R, Chen Q, Meerzaman D, et al. Scalable generation of pure CD103+ cDC1 from iDC1 cultures. Myeloid Cells. 2026;1:202613. https://doi.org/10.70401/mc.2026.0012

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