{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T13:49:52Z","timestamp":1790430592272,"version":"4.1.0"},"reference-count":36,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"4","license":[{"start":{"date-parts":[[2021,12,12]],"date-time":"2021-12-12T00:00:00Z","timestamp":1639267200000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":["Hepatology"],"published-print":{"date-parts":[[2022,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background and Aims<\/jats:title>\n            <jats:p>Pluripotent stem cell\u2013derived hepatocytes differentiated in monolayer culture are known to have more fetal than adult hepatocyte characteristics. If numerous studies tend to show that this immature phenotype might not necessarily be an obstacle to their use in transplantation, other applications such as drug screening, toxicological studies, or bioartificial livers are reliant on hepatocyte functionality and require full differentiation of hepatocytes. New technologies have been used to improve the differentiation process in recent years, usually evaluated by measuring the albumin production and CYP450 activity. Here we used the complex production and most importantly the activity of the coagulation factor IX (FIX) produced by mature hepatocytes to assess the differentiation of hemophilia B (HB) patient\u2019s induced pluripotent stem cells (iPSCs) in both monolayer culture and organoids.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Approach and Results<\/jats:title>\n            <jats:p>Indeed, HB is an X\u2010linked monogenic disease due to an impaired activity of FIX synthesized by hepatocytes in the liver. We have developed an in vitro model of HB hepatocytes using iPSCs generated from fibroblasts of a severe HB patient. We used CRISPR\/Cas9 technology to target the genomic insertion of a coagulation factor 9 minigene bearing the Padua mutation to enhance FIX activity. Noncorrected and corrected iPSCs were differentiated into hepatocytes under both two\u2010dimensional and three\u2010dimensional differentiation protocols and deciphered the production of active FIX in vitro. Finally, we assessed the therapeutic efficacy of this approach in vivo using a mouse model of HB.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>Functional FIX, whose post\u2010translational modifications only occur in fully mature hepatocytes, was only produced in corrected iPSCs differentiated in organoids. Immunohistochemistry analyses of mouse livers indicated a good cell engraftment, and the FIX activity detected in the plasma of transplanted animals confirmed rescue of the bleeding phenotype.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1002\/hep.32211","type":"journal-article","created":{"date-parts":[[2021,10,23]],"date-time":"2021-10-23T03:25:09Z","timestamp":1634959509000},"page":"866-880","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":20,"title":["In vitro recovery of FIX clotting activity as a marker of highly functional hepatocytes in a hemophilia B iPSC model"],"prefix":"10.1097","volume":"75","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8938-8405","authenticated-orcid":false,"given":"El\u00e9anor","family":"Luce","sequence":"first","affiliation":[{"name":"INSERM Universit\u00e9 Paris\u2010Saclay,Unit\u00e9 Mixte de Recherche 1193,Villejuif,France"},{"name":"F\u00e9deration Hospitalo\u2010Universitaire H\u00e9patinov, H\u00f4pital Paul Brousse,Villejuif,France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1201-4904","authenticated-orcid":false,"given":"Clara","family":"Steichen","sequence":"additional","affiliation":[{"name":"INSERM Universit\u00e9 Paris\u2010Saclay,Unit\u00e9 Mixte de Recherche 1193,Villejuif,France"},{"name":"F\u00e9deration Hospitalo\u2010Universitaire H\u00e9patinov, H\u00f4pital Paul Brousse,Villejuif,France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Micka\u00ebl","family":"Allouche","sequence":"additional","affiliation":[{"name":"INSERM Universit\u00e9 Paris\u2010Saclay,Unit\u00e9 Mixte de Recherche 1193,Villejuif,France"},{"name":"F\u00e9deration Hospitalo\u2010Universitaire H\u00e9patinov, H\u00f4pital Paul Brousse,Villejuif,France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5610-973X","authenticated-orcid":false,"given":"Antonietta","family":"Messina","sequence":"additional","affiliation":[{"name":"INSERM Universit\u00e9 Paris\u2010Saclay,Unit\u00e9 Mixte de Recherche 1193,Villejuif,France"},{"name":"F\u00e9deration Hospitalo\u2010Universitaire H\u00e9patinov, H\u00f4pital Paul Brousse,Villejuif,France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean\u2010Marie","family":"Heslan","sequence":"additional","affiliation":[{"name":"INSERM Unit\u00e9 Mixte de Recherche 1064,CHU H\u00f4tel Dieu,Nantes,France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6391-5839","authenticated-orcid":false,"given":"Thierry","family":"Lambert","sequence":"additional","affiliation":[{"name":"Centre de R\u00e9f\u00e9rence pour le Traitement des H\u00e9mophiles,H\u00f4pital de Bic\u00eatre,France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne","family":"Weber","sequence":"additional","affiliation":[{"name":"INSERM Universit\u00e9 Paris\u2010Saclay,Unit\u00e9 Mixte de Recherche 1193,Villejuif,France"},{"name":"F\u00e9deration Hospitalo\u2010Universitaire H\u00e9patinov, H\u00f4pital Paul Brousse,Villejuif,France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tuan Huy","family":"Nguyen","sequence":"additional","affiliation":[{"name":"INSERM Unit\u00e9 Mixte de Recherche 1064,CHU H\u00f4tel Dieu,Nantes,France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9080-6336","authenticated-orcid":false,"given":"Olivier","family":"Christophe","sequence":"additional","affiliation":[{"name":"INSERM Unit\u00e9 Mixte de Recherche 1176,H\u00f4pital de Bic\u00eatre,Kremlin\u2010Bic\u00eatre,France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9326-4413","authenticated-orcid":false,"given":"Anne","family":"Dubart\u2010Kupperschmitt","sequence":"additional","affiliation":[{"name":"INSERM Universit\u00e9 Paris\u2010Saclay,Unit\u00e9 Mixte de Recherche 1193,Villejuif,France"},{"name":"F\u00e9deration Hospitalo\u2010Universitaire H\u00e9patinov, H\u00f4pital Paul Brousse,Villejuif,France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","published-online":{"date-parts":[[2021,12,12]]},"reference":[{"key":"hep32211-bib-0001-20250824","article-title":"Novel treatments for hemophilia through rebalancing of the coagulation cascade","volume":"68","year":"2021","journal-title":"Pediatr Blood Cancer"},{"key":"hep32211-bib-0002-20250824","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1182\/blood-2018-07-820720","article-title":"Update on clinical gene therapy for 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