Impact of cell types and culture methods on the functionality of in vitro liver systems – A review of cell systems for hepatotoxicity assessment

Abstract

Xenobiotic safety assessment is an area that impacts a multitude of different industry sectors such as medicinal drugs, agrochemicals, industrial chemicals, cosmetics and environmental contaminants. As such there are a number of well-developed in vitro, in vivo and in silico approaches to evaluate their properties and potential impact on the environment and to humans. Additionally, there is the continual investment in multidisciplinary scientists to explore non-animal surrogate technologies to predict specific toxicological outcomes and to improve our understanding of the biological processes regarding the toxic potential of xenobiotics. Here we provide a concise, critical evaluation of a number of in vitro systems utilised to assess the hepatotoxic potential of xenobiotics.

Hepato(Geno)Toxicity Assessment of Nanoparticles in a HepG2 Liver Spheroid Model

“…The presence of non-parenchymal cells can possibly explain the higher response to the NPs in the InSphero model compared to the HepG2 spheroids. The HepG2 spheroids show relatively high metabolic capacity and appear to be a good advanced in vitro model for the liver [21][22][23][24]. The commercial primary cell InSphero co-culture model is more complex than the HepG2 spheroids.…”

“…However, when comparing in vitro cell culture models in standard two-dimensional (2D) monolayers with complex organs, the cell lines in 2D culture display a limited hepatocytic functionality [21].The liver-like functionality of the human hepatocellular carcinoma cell line HepG2 is enhanced when the cells are cultured in a three-dimensional (3D) arrangement. This increases the cell-to-cell contacts and intercellular communication [22] and changes the protein expression and metabolic status of the cells [21][22][23]. HepG2 cells in 3D cultures show upregulation of genes involved in liver-specific xenobiotic and lipid metabolism, whereas genes related to the extracellular matrix, cytoskeleton and cell adhesion have higher expression in 2D cultures [22,24].The use of spheroids as 3D cultures in hepatotoxicity assessment is an increasing field of interest, and HepG2 spheroids, prepared with and without using scaffolds, have been applied for toxicity experiments with both NPs [6,25,26] and chemicals [27,28].…”

“…The liver-like functionality of the human hepatocellular carcinoma cell line HepG2 is enhanced when the cells are cultured in a three-dimensional (3D) arrangement. This increases the cell-to-cell contacts and intercellular communication [22] and changes the protein expression and metabolic status of the cells [21][22][23]. HepG2 cells in 3D cultures show upregulation of genes involved in liver-specific xenobiotic and lipid metabolism, whereas genes related to the extracellular matrix, cytoskeleton and cell adhesion have higher expression in 2D cultures [22,24].…”

Gadolinium labelled nanoliposomes as the platform for MRI theranostics: in vitro safety study in liver cells and macrophages

“…HepaRG cells differentiate into hepatocyte-like colonies surrounded by biliary epithelial-like cells 19 and represent a liver model closest to primary hepatocytes. Compared to commonly used hepatic cell lines, HepaRG have better liver-specific functionality, such as CYP and nuclear receptor expressions 29 , and were shown to be a useful model for drug-induced human hepatotoxicity studies [30][31][32] . In this study, the HepaRG cells were used to screen possible in vitro toxicity of Gd-lip.…”

Type of endothelial cells affects HepaRG cell acetaminophen metabolism in both 2D and 3D porous scaffold cultures

“…The need for developing in vitro liver models mimicking native liver tissue is well understood (Kyffin et al, 2018). The frequently used HepG2 cell line to study the general cytotoxic potential of drugs expresses very low levels of phase I drug-metabolizing enzymes such as cytochrome P450s (Gerets et al, 2012;Takahashi et al, 2015).…”