Correspondence Address: Prof. Mehmet Ozturk, Faculty of Medicine, Izmir Tinaztepe University, Izmir 35400, Turkey. E-mail: firstname.lastname@example.org
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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 photos 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 the 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.
Conclusion: We describedexperimental conditions to obtain organoid-like structures from five different HCC cell lines representing hepatoblast-like and mesenchymal-like subgroups. These models are useful as an alternative to monolayer cultures to study phenotypic features of HCC cells. Our detailed analysis of biomarker expression in five different organoid-like structures provide convincing evidence for highly specific phenotypic features of these cell lines although they share some common or subtype-restricted features also.
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:[Accept]. http://dx.doi.org/10.20517/2394-5079.2021.43