Long-Term Adult Feline Liver Organoid Cultures for Disease Modeling of Hepatic Steatosis

Abstract

SummaryHepatic steatosis is a highly prevalent liver disease, yet research is hampered by the lack of tractable cellular and animal models. Steatosis also occurs in cats, where it can cause severe hepatic failure. Previous studies demonstrate the potential of liver organoids for modeling genetic diseases. To examine the possibility of using organoids to model steatosis, we established a long-term feline liver organoid culture with adult liver stem cell characteristics and differentiation potential toward hepatocyte-like cells. Next, organoids from mouse, human, dog, and cat liver were provided with fatty acids. Lipid accumulation was observed in all organoids and interestingly, feline liver organoids accumulated more lipid droplets than human organoids. Finally, we demonstrate effects of interference with β-oxidation on lipid accumulation in feline liver organoids. In conclusion, feline liver organoids can be successfully cultured and display a predisposition for lipid accumulation, making them an interesting model in hepatic steatosis research.

Organoids are promising tools for species‐specific in vitro toxicological studies

“…In each of these stages, different markers of hepatocyte maturity are expressed (Roskams, Cassiman, De Vos, & Libbrecht, 2004;Roskams, Libbrecht, & Desmet, 2003). Nowadays, for animal species (dog, cat), liver organoids are obtained using ASCs directly isolated from the liver tissue (Kruitwagen et al, 2017;Nantasanti et al, 2015), but not using iPSC-derived hepatocyte progenitors. Nowadays, for animal species (dog, cat), liver organoids are obtained using ASCs directly isolated from the liver tissue (Kruitwagen et al, 2017;Nantasanti et al, 2015), but not using iPSC-derived hepatocyte progenitors.…”

“…Spherical structures are formed by biliary duct fragments after digestion but also by undigested samples from fine-needle aspirate seeded in ECM (Matrigel) (Kruitwagen et al, 2017). Spherical structures are formed by biliary duct fragments after digestion but also by undigested samples from fine-needle aspirate seeded in ECM (Matrigel) (Kruitwagen et al, 2017).…”

“…Spherical structures are formed by biliary duct fragments after digestion but also by undigested samples from fine-needle aspirate seeded in ECM (Matrigel) (Kruitwagen et al, 2017). Kruitwagen and colleagues (Kruitwagen et al, 2017) described a method to generate and culture 3D liver organoids, which is, to our knowledge, the only available primary nontransformed long-term cell culture system for the feline species. The liver ASC populations play an important role in feline organoid generation.…”

“…Feline liver organoids are constituted of cells characterized by a long genetic stability and, within their structure, as for other types of organoids, cells at different maturation stages can be found (Sato et al, 2009). The feline liver organoids displayed a single-layered cubical epithelium and were positive for an E-cadherin marker (an epithelial marker) (Kruitwagen et al, 2017). Kruitwagen and colleagues (Kruitwagen et al, 2017) described a method to generate and culture 3D liver organoids, which is, to our knowledge, the only available primary nontransformed long-term cell culture system for the feline species.…”

“…The liver ASC populations play an important role in feline organoid generation. Kruitwagen and colleagues (Kruitwagen et al, 2017) have succeeded in improving the hepatocyte function (upregulation of aminotransferase levels, albumin secretion and CYP450 activity) and intermediate hepatocyte phenotype during organoid differentiation. The feline liver organoids displayed a single-layered cubical epithelium and were positive for an E-cadherin marker (an epithelial marker) (Kruitwagen et al, 2017).…”