{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T20:51:43Z","timestamp":1776286303184,"version":"3.50.1"},"reference-count":13,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T00:00:00Z","timestamp":1768780800000},"content-version":"vor","delay-in-days":18,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,1]]},"abstract":"Abstract<\/jats:title>\n In vitro modeling of human liver function is essential for assessing drug\u2010induced toxicity, particularly in the context of metabolic dysfunction\u2013associated steatotic liver disease (MASLD). However, standard liver organoid systems often lack xenobiotic\u2010metabolizing enzyme activity and fail to replicate the pathological features of MASLD. Here, we present a method for generating functional, multi cell\u2013type human liver organoids (HML) organoids from HepaRG cells, primary human macrophages, and LX\u20102 hepatic stellate cells. These three\u2010dimensional (3D) organoids are cultured under defined conditions that support key hepatic functions. To mimic MASLD, we expose HML organoids to a mixture of stearic and oleic acids for 9 days, inducing steatosis and fibrogenic responses characteristic of the disease. These MASLD\u2010HML organoids retain liver\u2010specific functions and express key fibrosis markers. We further demonstrate the usefulness of this method to produce standardized organoids useful for the evaluation of drug\u2010induced liver injury (DILI) through IC50 and benchmark dose calculations, particularly in the context of MASLD. Together, HML and MASLD\u2010HML organoids provide a robust model for studying drug metabolism, toxicity, and adverse drug reactions in healthy and diseased liver states. \u00a9 2026 Wiley Periodicals LLC.<\/jats:p>\n \n Basic Protocol 1<\/jats:bold>\n : Culture of HepaRG cells\n <\/jats:p>\n \n Basic Protocol 2<\/jats:bold>\n : Culture of LX\u20102 cells\n <\/jats:p>\n \n Basic Protocol 3<\/jats:bold>\n : Culture of primary human unstimulated M1 macrophages\n <\/jats:p>\n \n Basic Protocol 4<\/jats:bold>\n : Organoid seeding procedure 1 in agarose molds\n <\/jats:p>\n \n Basic Protocol 5<\/jats:bold>\n : Organoid seeding procedure 2 in ultra\u2010low attachment 96\u2010well plate\n <\/jats:p>\n \n Alternate Protocol<\/jats:bold>\n : Organoid seeding procedure from cryopreserved cells\n <\/jats:p>\n \n Basic Protocol 6<\/jats:bold>\n : Preparation of the fatty acid mixture\n <\/jats:p>\n \n Basic Protocol 7<\/jats:bold>\n : MASLD induction\n <\/jats:p>","DOI":"10.1002\/cpz1.70294","type":"journal-article","created":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T10:39:51Z","timestamp":1768819191000},"source":"Crossref","is-referenced-by-count":1,"title":["Multi\u2010Cellular Human Liver Organoids for Modeling Metabolic Dysfunction\u2013Associated Steatotic Liver Disease and Drug\u2010Induced Liver Injury"],"prefix":"10.1002","volume":"6","author":[{"given":"Maela","family":"Duclos","sequence":"first","affiliation":[{"name":"Institut NuMeCan (Nutrition, Metabolisms and Cancer), INSERM 1317, UMR INRAE 1341 Universit\u00e9 de Rennes Rennes France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7398-3816","authenticated-orcid":false,"given":"Zeineb","family":"Marzougui","sequence":"additional","affiliation":[{"name":"Toxicology of Contaminants Unit, Foug\u00e8res Laboratory, French Agency for Food Environmental and Occupational Health & Safety (ANSES) Foug\u00e8res France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mariam","family":"Saleh","sequence":"additional","affiliation":[{"name":"Toxicology of Contaminants Unit, Foug\u00e8res Laboratory, French Agency for Food Environmental and Occupational Health & Safety (ANSES) Foug\u00e8res France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anne","family":"Corlu","sequence":"additional","affiliation":[{"name":"Institut NuMeCan (Nutrition, Metabolisms and Cancer), INSERM 1317, UMR INRAE 1341 Universit\u00e9 de Rennes Rennes France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julie","family":"Massart","sequence":"additional","affiliation":[{"name":"Institut NuMeCan (Nutrition, Metabolisms and Cancer), INSERM 1317, UMR INRAE 1341 Universit\u00e9 de Rennes Rennes France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7417-4690","authenticated-orcid":false,"given":"Pierre\u2010Jean","family":"Ferron","sequence":"additional","affiliation":[{"name":"Toxicology of Contaminants Unit, Foug\u00e8res Laboratory, French Agency for Food Environmental and Occupational Health & Safety (ANSES) Foug\u00e8res France"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2026,1,19]]},"reference":[{"key":"e_1_2_13_2_1","doi-asserted-by":"publisher","DOI":"10.1124\/dmd.105.006759"},{"key":"e_1_2_13_3_1","doi-asserted-by":"publisher","DOI":"10.1055\/a\u20102404\u20107973"},{"key":"e_1_2_13_4_1","doi-asserted-by":"publisher","DOI":"10.3390\/biomedicines13112722"},{"key":"e_1_2_13_5_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13318\u2010018\u20100533\u20103"},{"key":"e_1_2_13_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tiv.2023.105728"},{"key":"e_1_2_13_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/hep.21536"},{"key":"e_1_2_13_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.232137699"},{"key":"e_1_2_13_9_1","doi-asserted-by":"publisher","DOI":"10.1124\/dmd.109.031831"},{"key":"e_1_2_13_10_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms221910214"},{"key":"e_1_2_13_11_1","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v23.i47.8263"},{"key":"e_1_2_13_12_1","doi-asserted-by":"publisher","DOI":"10.1097\/HEP.0000000000000520"},{"key":"e_1_2_13_13_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms222312782"},{"key":"e_1_2_13_14_1","doi-asserted-by":"publisher","DOI":"10.1248\/bpb.b17\u201000913"}],"container-title":["Current 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