{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T15:22:35Z","timestamp":1791386555155,"version":"4.3.3"},"reference-count":61,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2019,4,2]],"date-time":"2019-04-02T00:00:00Z","timestamp":1554163200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004359","name":"Vetenskapsr\u00e5det","doi-asserted-by":"publisher","award":["2016-01527"],"award-info":[{"award-number":["2016-01527"]}],"id":[{"id":"10.13039\/501100004359","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003793","name":"Hj\u00e4rt-Lungfonden","doi-asserted-by":"publisher","award":["20120533"],"award-info":[{"award-number":["20120533"]}],"id":[{"id":"10.13039\/501100003793","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the Swedish state under the agreement between the Swedish government and the county councils, the ALF-agreement","award":["SU 2018-04276"],"award-info":[{"award-number":["SU 2018-04276"]}]},{"name":"Novonordisk Foundation Grant for Excellence in Endocrinology","award":["Excellence Project, 9321-430"],"award-info":[{"award-number":["Excellence Project, 9321-430"]}]},{"DOI":"10.13039\/501100008546","name":"Swedish Diabetes Foundation","doi-asserted-by":"publisher","award":["DIA 2017-205"],"award-info":[{"award-number":["DIA 2017-205"]}],"id":[{"id":"10.13039\/501100008546","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Research grant from AstraZeneca","award":["Echo Project, 10033852"],"award-info":[{"award-number":["Echo Project, 10033852"]}]},{"name":"Wallenberg Academy Fellows from the Knut and Alice Wallenberg Foundation","award":["KAW 2017.0203"],"award-info":[{"award-number":["KAW 2017.0203"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Non-alcoholic fatty liver disease (NAFLD) is the most common liver disorder in western countries. Despite the high prevalence of NAFLD, the underlying biology of the disease progression is not clear, and there are no approved drugs to treat non-alcoholic steatohepatitis (NASH), the most advanced form of the disease. Thus, there is an urgent need for developing advanced in vitro human cellular systems to study disease mechanisms and drug responses. We attempted to create an organoid system genetically predisposed to NAFLD and to induce steatosis and fibrosis in it by adding free fatty acids. We used multilineage 3D spheroids composed by hepatocytes (HepG2) and hepatic stellate cells (LX-2) with a physiological ratio (24:1). HepG2 and LX-2 cells are homozygotes for the PNPLA3 I148M sequence variant, the strongest genetic determinant of NAFLD. We demonstrate that hepatic stellate cells facilitate the compactness of 3D spheroids. Then, we show that the spheroids develop accumulations of fat and collagen upon exposure to free fatty acids. Finally, this accumulation was rescued by incubating spheroids with liraglutide or elafibranor, drugs that are in clinical trials for the treatment of NASH. In conclusion, we have established a simple, easy to handle, in vitro model of genetically induced NAFLD consisting of multilineage 3D spheroids. This tool may be used to understand molecular mechanisms involved in the early stages of fibrogenesis induced by lipid accumulation. Moreover, it may be used to identify new compounds to treat NASH using high-throughput drug screening.<\/jats:p>","DOI":"10.3390\/ijms20071629","type":"journal-article","created":{"date-parts":[[2019,4,3]],"date-time":"2019-04-03T03:39:28Z","timestamp":1554262768000},"page":"1629","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":137,"title":["Human Multilineage 3D Spheroids as a Model of Liver Steatosis and Fibrosis"],"prefix":"10.3390","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2478-225X","authenticated-orcid":false,"given":"Piero","family":"Pingitore","sequence":"first","affiliation":[{"name":"Department of Molecular and Clinical Medicine, University of Gothenburg, SE-413 45 Gothenburg, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kavitha","family":"Sasidharan","sequence":"additional","affiliation":[{"name":"Department of Molecular and Clinical Medicine, University of Gothenburg, SE-413 45 Gothenburg, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0403-9266","authenticated-orcid":false,"given":"Matias","family":"Ekstrand","sequence":"additional","affiliation":[{"name":"Department of Molecular and Clinical Medicine, University of Gothenburg, SE-413 45 Gothenburg, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sebastian","family":"Prill","sequence":"additional","affiliation":[{"name":"Cardiovascular, Renal and Metabolism, IMED Biotech Unit, AstraZeneca, SE-431 83 Gothenburg, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4158-8018","authenticated-orcid":false,"given":"Daniel","family":"Lind\u00e9n","sequence":"additional","affiliation":[{"name":"Cardiovascular, Renal and Metabolism, IMED Biotech Unit, AstraZeneca, SE-431 83 Gothenburg, Sweden"},{"name":"Division of Endocrinology, Department of Neuroscience and Physiology, Sahlgrenska Academy, University of Gothenburg, SE-405 30 Gothenburg, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefano","family":"Romeo","sequence":"additional","affiliation":[{"name":"Department of Molecular and Clinical Medicine, University of Gothenburg, SE-413 45 Gothenburg, Sweden"},{"name":"Clinical Nutrition Unit, Department of Medical and Surgical Sciences, Magna Graecia University, 88100 Catanzaro, Italy"},{"name":"Cardiology Department, Sahlgrenska University Hospital, SE-413 45 Gothenburg, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,4,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1016\/j.jhep.2012.12.005","article-title":"The burden of liver disease in Europe: A review of available epidemiological data","volume":"58","author":"Blachier","year":"2013","journal-title":"J. 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