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Bioprinting and most alternative technologies rely on animal-derived materials, which compromises the intent of animal welfare and results in the generation of chimeric systems of limited value. The current study therefore presents the first bioprinted liver model that is entirely void of animal-derived constituents. Initially, HuH-7 cells underwent adaptation to a chemically defined medium (CDM). The adapted cells exhibited high survival rates (85\u201392%) after cryopreservation in chemically defined freezing media, comparable to those preserved in standard medium (86\u201392%). Xeno-free bioink for 3D bioprinting yielded liver models with high relative cell viability (97\u2013101%), akin to a Matrigel-based liver model (83\u2013102%) after 15 days of culture. The established xeno-free model was used for toxicity testing of a marine biotoxin, okadaic acid (OA). In 2D culture, OA toxicity was virtually identical for cells cultured under standard conditions and in CDM. In the xeno-free bioprinted liver model, 3-fold higher concentrations of OA than in the respective monolayer culture were needed to induce cytotoxicity. In conclusion, this study describes for the first time the development of a xeno-free 3D bioprinted liver model and its applicability for research purposes.<\/jats:p>","DOI":"10.3390\/ijms25031811","type":"journal-article","created":{"date-parts":[[2024,2,2]],"date-time":"2024-02-02T09:24:08Z","timestamp":1706865848000},"page":"1811","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Xeno-Free 3D Bioprinted Liver Model for Hepatotoxicity Assessment"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0001-8222","authenticated-orcid":false,"given":"Ahmed S. M.","family":"Ali","sequence":"first","affiliation":[{"name":"Department of Applied Biochemistry, Institute of Biotechnology, Technische Universit\u00e4t Berlin, TIB 4\/3-2, Gustav-Meyer-Allee 25, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johanna","family":"Berg","sequence":"additional","affiliation":[{"name":"Department of Applied Biochemistry, Institute of Biotechnology, Technische Universit\u00e4t Berlin, TIB 4\/3-2, Gustav-Meyer-Allee 25, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Viola","family":"Roehrs","sequence":"additional","affiliation":[{"name":"Department of Applied Biochemistry, Institute of Biotechnology, Technische Universit\u00e4t Berlin, TIB 4\/3-2, Gustav-Meyer-Allee 25, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5446-7376","authenticated-orcid":false,"given":"Dongwei","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Applied Biochemistry, Institute of Biotechnology, Technische Universit\u00e4t Berlin, TIB 4\/3-2, Gustav-Meyer-Allee 25, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johannes","family":"Hackethal","sequence":"additional","affiliation":[{"name":"THT Biomaterials, 1030 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Albert","family":"Braeuning","sequence":"additional","affiliation":[{"name":"Department Food Safety, German Federal Institute for Risk Assessment (BfR), 10589 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Woelken","sequence":"additional","affiliation":[{"name":"Department of Food Biotechnology and Food Process Engineering, Technische Universit\u00e4t Berlin, 14195 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4696-1128","authenticated-orcid":false,"given":"Cornelia","family":"Rauh","sequence":"additional","affiliation":[{"name":"Department of Food Biotechnology and Food Process Engineering, Technische Universit\u00e4t Berlin, 14195 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1469-0052","authenticated-orcid":false,"given":"Jens","family":"Kurreck","sequence":"additional","affiliation":[{"name":"Department of Applied Biochemistry, Institute of Biotechnology, Technische Universit\u00e4t Berlin, TIB 4\/3-2, Gustav-Meyer-Allee 25, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Tabernilla, A., Dos Santos Rodrigues, B., Pieters, A., Caufriez, A., Leroy, K., Van Campenhout, R., Cooreman, A., Gomes, A.R., Arnesdotter, E., and Gijbels, E. 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