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However, culturing hepatocytes as three\u2010dimensional (3D) spheroids has been shown to result in improved liver\u2010specific functions (e.g., metabolic capacity) by better mimicking the in vivo environment. This approach may lead to more reliable detection of drug\u2010induced liver injury (DILI) in the early phase of drug discovery, preventing post\u2010marketing drug withdrawals. Here, we investigated the cultivation of TAMH as 3D spheroids, characterizing them with optical and transmission electron microscopy as well as analyzing their gene expression at mRNA level (especially drug\u2010metabolizing enzymes) compared to TAMH monolayer. In addition, comparisons were made with spheroids grown from the human hepatoblastoma cell line HepG2, another current spheroid model. The results indicate that TAMH spheroids express hepatic structures and show elevated levels of some of the key phase I and II drug\u2010metabolizing enzymes, in contrast to TAMH monolayer. The in vitro hepatotoxic potencies of the drugs acetaminophen and flupirtine maleate were found to be very similar between TAMH spheroidal and the monolayer cultures. Both the advantages and disadvantages of TAMH spheroids as an in vitro hepatotoxicity model compared to monolayer model are discussed.<\/jats:p>","DOI":"10.1002\/jat.4538","type":"journal-article","created":{"date-parts":[[2023,9,1]],"date-time":"2023-09-01T06:38:13Z","timestamp":1693550293000},"page":"272-286","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Investigation of TGF\u2010<i>\u03b1<\/i>\u2010overexpressing mouse hepatocytes (TAMH) cultured as spheroids for use in hepatotoxicity screening of drug candidates"],"prefix":"10.1002","volume":"44","author":[{"given":"Kristin","family":"Beirow","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical\/Medicinal Chemistry, Institute of Pharmacy University of Greifswald  Greifswald Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christian","family":"Schmidt","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Biotechnology Institute of Pharmacy University of Greifswald  Greifswald Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Britta","family":"J\u00fcrgen","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Biotechnology Institute of Pharmacy University of Greifswald  Greifswald Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rabea","family":"Schl\u00fcter","sequence":"additional","affiliation":[{"name":"Imaging Center of the Department of Biology University of Greifswald  Greifswald Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas","family":"Schweder","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Biotechnology Institute of Pharmacy University of Greifswald  Greifswald Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9589-8241","authenticated-orcid":false,"given":"Patrick J.","family":"Bednarski","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical\/Medicinal Chemistry, Institute of Pharmacy University of Greifswald  Greifswald Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2023,9]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-04-0496"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1517\/17425255.4.11.1415"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1254\/jphs.13135FP"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/cmdc.202300145"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1093\/toxsci\/kfx289"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep25187"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1124\/dmd.116.074369"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.4155\/fmc-2018-0350"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.metabol.2006.01.006"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yebeh.2019.106580"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/tx7001349"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmedt.2020.611913"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1155\/2016\/4780872"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1080\/0049825031000076168"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1089\/adt.2014.573"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-41273-3"},{"key":"e_1_2_7_18_1","unstructured":"European Medicines Agency. 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