{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T06:53:52Z","timestamp":1792652032496,"version":"4.5.0"},"reference-count":52,"publisher":"National Academy of Sciences","issue":"42","license":[{"start":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T00:00:00Z","timestamp":1760486400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003246","name":"Nederlandse Organisatie voor Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["024.003.013"],"award-info":[{"award-number":["024.003.013"]}],"id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003246","name":"Nederlandse Organisatie voor Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["20941"],"award-info":[{"award-number":["20941"]}],"id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100021821","name":"Oncode Institute","doi-asserted-by":"publisher","award":["01"],"award-info":[{"award-number":["01"]}],"id":[{"id":"10.13039\/501100021821","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001826","name":"ZonMw","doi-asserted-by":"publisher","award":["01142052310003"],"award-info":[{"award-number":["01142052310003"]}],"id":[{"id":"10.13039\/501100001826","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2025,10,21]]},"abstract":"<jats:p>\n                    Tissue stem cell (TSC)-derived epithelial organoids are typically cultured in Matrigel [T. Sato\n                    <jats:italic toggle=\"yes\">et al.<\/jats:italic>\n                    ,\n                    <jats:italic toggle=\"yes\">Nature<\/jats:italic>\n                    <jats:bold>459<\/jats:bold>\n                    , 262\u2013265 (2009)], an extracellular matrix-like hydrogel produced from Engelbreth\u2013Holm\u2013Swarm sarcoma cells. This tumor is grown in the mouse abdomen [R. W. Orkin\n                    <jats:italic toggle=\"yes\">et al.<\/jats:italic>\n                    ,\n                    <jats:italic toggle=\"yes\">J. Exp. Med.<\/jats:italic>\n                    <jats:bold>145<\/jats:bold>\n                    , 204\u2013220 (1977)]. Previously, we demonstrated that the\n                    <jats:italic toggle=\"yes\">\n                      <jats:italic toggle=\"yes\">Yersinia<\/jats:italic>\n                    <\/jats:italic>\n                    membrane protein Invasin, coated on transwells, replaces Matrigel by activating \u03b21-integrins, allowing long-term expansion of primary epithelial cells as 2D organoid sheets [J. J. A. P. M. Wijnakker\n                    <jats:italic toggle=\"yes\">et al.<\/jats:italic>\n                    ,\n                    <jats:italic toggle=\"yes\">Proc. Natl. Acad. Sci. U.S.A.<\/jats:italic>\n                    <jats:bold>122<\/jats:bold>\n                    , e2420595121 (2025)]. Here, we functionalize a synthetic polyisocyanide (PIC) hydrogel with the integrin-activating domain of Invasin (INV). PIC hydrogels are soluble at 4 \u00b0C and form a gel at 37 \u00b0C [P. H. J. Kouwer\n                    <jats:italic toggle=\"yes\">et al.<\/jats:italic>\n                    ,\n                    <jats:italic toggle=\"yes\">Nature<\/jats:italic>\n                    <jats:bold>493<\/jats:bold>\n                    , 651\u2013655 (2013)]. When INV is covalently linked to PIC, the resulting hydrogel supports multipassage 3D growth of human intestinal and airway organoids. Self-renewal, polarization, and differentiation are maintained. The 3D swelling assay for cystic fibrosis drug testing (S. F. Boj\n                    <jats:italic toggle=\"yes\">et al.<\/jats:italic>\n                    ,\n                    <jats:italic toggle=\"yes\">J. Vis. Exp.<\/jats:italic>\n                    (2017), 10.3791\/55159] was validated using PIC-INV. With PIC-INV hydrogels, we establish a fully defined and animal-free system for 3D TSC-derived organoid culture.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2507500122","type":"journal-article","created":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T17:39:33Z","timestamp":1760549973000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":16,"title":["Invasin-functionalized PIC hydrogels enable long-term 3D culture of epithelial organoids"],"prefix":"10.1073","volume":"122","author":[{"given":"Joost J. A. P. M.","family":"Wijnakker","sequence":"first","affiliation":[{"name":"Oncode Institute, Hubrecht Institute, Royal Netherlands Academy of Arts and Science (KNAW)","place":["Utrecht, The Netherlands"]},{"name":"University Medical Centre","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sangho","family":"Lim","sequence":"additional","affiliation":[{"name":"Oncode Institute, Hubrecht Institute, Royal Netherlands Academy of Arts and Science (KNAW)","place":["Utrecht, The Netherlands"]},{"name":"University Medical Centre","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robin","family":"Schreurs","sequence":"additional","affiliation":[{"name":"Oncode Institute, Hubrecht Institute, Royal Netherlands Academy of Arts and Science (KNAW)","place":["Utrecht, The Netherlands"]},{"name":"University Medical Centre","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u00e3o Ferreira","family":"Faria","sequence":"additional","affiliation":[{"name":"Institute for Molecules and Materials, Radboud University","place":["Nijmegen 6525 AJ, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeroen","family":"Korving","sequence":"additional","affiliation":[{"name":"Oncode Institute, Hubrecht Institute, Royal Netherlands Academy of Arts and Science (KNAW)","place":["Utrecht, The Netherlands"]},{"name":"University Medical Centre","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harry","family":"Begthel","sequence":"additional","affiliation":[{"name":"Oncode Institute, Hubrecht Institute, Royal Netherlands Academy of Arts and Science (KNAW)","place":["Utrecht, The Netherlands"]},{"name":"University Medical Centre","place":["Utrecht, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kirti K.","family":"Iyer","sequence":"additional","affiliation":[{"name":"Institute for Molecules and Materials, Radboud University","place":["Nijmegen 6525 AJ, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul H. J.","family":"Kouwer","sequence":"additional","affiliation":[{"name":"Institute for Molecules and Materials, Radboud University","place":["Nijmegen 6525 AJ, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3077-5582","authenticated-orcid":false,"given":"Hans","family":"Clevers","sequence":"additional","affiliation":[{"name":"Oncode Institute, Hubrecht Institute, Royal Netherlands Academy of Arts and Science (KNAW)","place":["Utrecht, The Netherlands"]},{"name":"University Medical Centre","place":["Utrecht, The Netherlands"]},{"name":"Institute of Human Biology (IHB), Roche Pharma Research and Early Development, Roche Innovation Center","place":["Basel, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2025,10,15]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.cellbio.22.010305.104315"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2019.01.108"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1242\/dev.028423"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.145.1.204"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00350a005"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)83607-4"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.4161\/cam.21831"},{"key":"e_1_3_4_8_2","first-page":"2","article-title":"Architecture is the message: The role of extracellular matrix and 3-D structure in tissue-specific gene expression and breast cancer","volume":"16","author":"Bissell M. 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