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Here we demonstrate that precise modulation of Activin A concentrations within a dynamic 3D vertical-wheel bioreactor system enables the co-emergence and spatial organization of hepatic epithelium and septum transversum mesenchyme-like populations from human pluripotent stem cells. High Activin levels induce uniform definitive endoderm and enhanced hepatic maturation, whereas low levels promote epithelial tube\u2013like structures encased by mesenchymal cells, recapitulating early liver diverticulum architecture. This gradient-based approach autonomously generates parenchymal and non-parenchymal hepatic lineages, providing a scalable platform to model human liver organogenesis and engineer ventral foregut\u2013derived tissues.<\/jats:p>","DOI":"10.1101\/2025.11.27.690974","type":"posted-content","created":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T08:30:15Z","timestamp":1764664215000},"source":"Crossref","is-referenced-by-count":0,"title":["Gradient-Based Regulation of Activin A Directs Multilineage Liver Organoid Development from Human Pluripotent Stem Cells"],"prefix":"10.64898","author":[{"given":"Jo\u00e3o P.","family":"Cotovio","sequence":"first","affiliation":[{"name":"iBB - Institute for Bioengineering and Biosciences and Department of Bioengineering, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Lisbon, Portugal"},{"name":"Associate Laboratory i4HB Institute for Health and Bioeconomy, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Lisbon, Portugal"}]},{"given":"Alice C.","family":"Borges","sequence":"additional","affiliation":[{"name":"iBB - Institute for Bioengineering and Biosciences and Department of Bioengineering, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Lisbon, Portugal"},{"name":"Associate Laboratory i4HB Institute for Health and Bioeconomy, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Lisbon, Portugal"},{"name":"Gulbenkian Institute for Molecular Medicine (GIMM), Lisbon, Portugal"}]},{"given":"Diogo E.S.","family":"Nogueira","sequence":"additional","affiliation":[{"name":"iBB - Institute for Bioengineering and Biosciences and Department of Bioengineering, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Lisbon, Portugal"},{"name":"Associate Laboratory i4HB Institute for Health and Bioeconomy, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Lisbon, Portugal"}]},{"given":"Carlos A. V.","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"iBB - Institute for Bioengineering and Biosciences and Department of Bioengineering, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Lisbon, Portugal"},{"name":"Associate Laboratory i4HB Institute for Health and Bioeconomy, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Lisbon, Portugal"}]},{"given":"Joaquim M. 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