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A diverse population of stroma cells provides surface\u2010bound and soluble molecules that orchestrate the intrathymic maturation and selection of developing T cells. Forming an intricate 3D architecture, thymic epithelial cells (TEC) represent the most abundant and important constituent of the thymic stroma. Effective models for in and ex vivo use of adult TEC are still wanting, limiting the engineering of functional thymic organoids and the understanding of the development of a competent immune system. Here a 3D scaffold is developed based on decellularized thymic tissue capable of supporting in vitro and in vivo thymopoiesis by both fetal and adult TEC. For the first time, direct evidences of feasibility for sustained graft\u2010resident T\u2010cell development using adult TEC as input are provided. Moreover, the scaffold supports prolonged in vitro culture of adult TEC, with a retained expression of the master regulator Foxn1. The success of engineering a thymic scaffold that sustains adult TEC function provides unprecedented opportunities to investigate thymus development and physiology and to design and implement novel strategies for thymus replacement therapies.<\/jats:p>","DOI":"10.1002\/adfm.202010747","type":"journal-article","created":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T04:40:38Z","timestamp":1615524038000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":42,"title":["Thymus Extracellular Matrix\u2010Derived Scaffolds Support Graft\u2010Resident Thymopoiesis and Long\u2010Term In Vitro Culture of Adult Thymic Epithelial Cells"],"prefix":"10.1002","volume":"31","author":[{"given":"M. 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