Oz et al. Hepatoma Res 2021;7:60 DOI: 10.20517/2394-5079.2021.43 Hepatoma Research © The Author(s) 2021. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, sharing, adaptation, distribution and reproduction in any medium or format, for any purpose, even commercially, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. www.hrjournal.net Open Access Original Article 3D Organoid modelling of hepatoblast-like and mesenchymal-like hepatocellular carcinoma cell lines Ozden Oz 1,2,3 , Evin Iscan 1,2 , Tugce Batur 1,2 , Mehmet Ozturk 1,4 1 Izmir Biomedicine and Genome Center, Izmir 35000, Turkey. 2 Izmir International Biomedicine and Genome Institute, Dokuz Eylul University, Izmir 35000, Turkey. 3 Izmir Bozyaka Education and Research Hospital, University of Health Sciences, Izmir 35000, Turkey. 4 Faculty of Medicine, Izmir Tinaztepe University, Izmir 35400, Turkey Correspondence to: Prof. Mehmet Ozturk, Faculty of Medicine, Izmir Tinaztepe University, Izmir 35400, Turkey. E-mail: mehmet.ozturk@tinztepe.edu.tr How to cite this article: Oz O, Iscan E, Batur T, Ozturk M. 3D Organoid modelling of hepatoblast-like and mesenchymal-like hepatocellular carcinoma cell lines. Hepatoma Res 2021;7:60. https://dx.doi.org/10.20517/2394-5079.2021.43 Received: 31 Mar 2021 First Decision: 11 May 2021 Revised: 15 Jun 2021 Accepted: 13 Jul 2021 First online: 20 Jul 2021 Academic Editor: Guang-Wen Cao Copy Editor: Xi-Jun Chen Production Editor: Xi-Jun Chen Abstract Aim: We wished to establish 3D organoid-like hepatocellular carcinoma (HCC) models from HCC cell lines. Methods: Hep3B, Huh7, HepG2, SNU398, SNU449, and SNU475 cell lines were inoculated into Matrigel and grown up to 9 days in hepatocyte specific or standard RPMI media. Spheroid formation was followed by light microscopy. Matrigel scaffolds were immobilized and embedded in paraffin, and sections were subjected to H&E and immunohistochemical staining for different hepatobiliary biomarkers. Stained material was examined under light microscopy and micro photo were taken. Results: Organoid-like structures were obtained successfully from all selected cell lines except mesenchymal-like SNU475 cells. Hep3B, Huh7, and HepG2 cell lines from hepatoblast-like sub-group formed compact 3D colonies and showed hepatocyte-like morphology and staining with different hepatocyte lineage markers as well as hepatobiliary progenitor markers. SNU398 and SNU449 cell lines from mesenchymal-like group formed irregular and loose 3D colonies that expressed vimentin homogeneously, but also several epithelial and hepatocyte lineage markers. The pattern of biomarker expression was unique for each cell line tested. Such features, not observed in tested monolayer cultures were confirmed with single-cell derived Hep3B cells.