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Previous studies suggest that the thymic DC pool consists of plasmacytoid DC (pDC), XCR1+ DC1, and SIRP\u03b1+ DC2. Yet the precise origin, development, and homeostasis, particularly of DC2, remain unresolved. Using single-cell transcriptomics and lineage-defining mouse models, we identify nine major populations of thymic myeloid cells and describe their lineage identities. What was previously considered to be \u201cDC2\u201d is actually composed of four distinct cell lineages. Among these are monocyte-derived DCs (moDCs) and monocyte-derived macrophages (moMacs), which are dependent on thymic IFN to upregulate MHCII and CD11c. We further demonstrate that conventional DC2 undergo intrathymic maturation through CD40 signaling. Finally, amongst DC2, we identify a novel thymic population of CX3CR1+ transitional DC (tDC), which represents transendothelial DCs positioned near thymic microvessels. Together, these findings reveal the thymus as a niche for diverse, developmentally distinct myeloid cells and elucidate their specific requirements for development and maturation.<\/jats:p>","DOI":"10.1084\/jem.20250733","type":"journal-article","created":{"date-parts":[[2025,8,27]],"date-time":"2025-08-27T06:20:55Z","timestamp":1756275655000},"update-policy":"https:\/\/doi.org\/10.1084\/jem.crossmarkpolicy","source":"Crossref","is-referenced-by-count":7,"title":["Thymic myeloid cells are heterogenous and include a novel population of transitional dendritic cells"],"prefix":"10.1084","volume":"223","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3672-7629","authenticated-orcid":true,"given":"Matou\u0161","family":"Vobo\u0159il","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Center for Immunology, Department of Lab Medicine and Pathology, University of Minnesota Medical School 1 , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/024d6js02","id-type":"ROR","asserted-by":"publisher"}],"name":"Charles University 2 Department of Cell Biology, Faculty of Science, , Prague,","place":["Czech Republic"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5611-6967","authenticated-orcid":true,"given":"Fernando Bandeira","family":"Sulczewski","sequence":"additional","affiliation":[{"name":"University of San Diego School of Medicine 3 Department of Pharmacology, , San Diego, CA,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2045-8794","authenticated-orcid":true,"given":"Ryan J.","family":"Martinez","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Center for Immunology, Department of Lab Medicine and Pathology, University of Minnesota Medical School 1 , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6297-5941","authenticated-orcid":true,"given":"K. 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