The development of three-dimensional liver lobule-like platforms for evaluating hepatotoxicity
Korea Institute of Toxicology
To date, the various in vitro liver models such as three dimensional (3D) hepatocyte spheroids and artificial liver tissues have been investigated to evaluate hepatotoxicity by mimicking the in vivo liver environment. Although there are big advances in developing in vitro liver models, there are still limitations in providing multiple read-outs for hepatotoxicity from in vitro liver models. Here we develop platforms mimicking the hepatic lobule structure based on 3D scaffolds, to easily apply hepatotoxicity assays and evaluate the hepatotoxicity of different subtypes of liver cells. This 3D lobule-like platform provides inserted modules consisting of hepatocytes, Kupffer cells, and stellate cells embedded into 3D porous scaffold. Each cell module containing scaffolds can be inserted into the liver lobules-like structure for co-culture and may be easily detached from the structure to observe and evaluate hepatotoxicity. The surface of the 3D porous scaffold, made of polymethylasiloxane (PDMS), is modified by oxygen plasma to increase the hydrophilicity and matrigel is coated on the scaffold. The three different cells including HepG2 for hepatocytes, Lx-2 for stellate cells and Raw 264.7 for macrophages were cultured on 3D culture module and co-culture condition was optimized. Using this lobule-like platform allows for efficient hepatotoxicity testing and recovery of samples for analyzing the RNA or proteins from each cell type. This platform can be used to analyze the multiple-reads of toxicity from the co-cultured cells and to understand the toxic mechanisms underlying interactive effects among the parenchymal and non-parenchymal liver cells. This work was supported by a grant (NRF-2016M3A9C4953144) from the Ministry of Science, ICT, and Future Planning and a general research grant from the Korea Institute of Toxicology.