A novel differentiated HuH-7 cell model to examine bile acid metabolism, transport and cholestatic hepatotoxicity

• Published in: Scientific reports Vol. 12; no. 1; pp. 14333 - 14
• Main Authors: Saran, Chitra, Fu, Dong, Ho, Henry, Klein, Abigail, Fallon, John K., Honkakoski, Paavo, Brouwer, Kim L. R.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 22.08.2022; Nature Publishing Group; Nature Portfolio
• Subjects: 631/154/570; 631/1647/767; 631/80/85; 631/92/577; 692/4020/4021; Acyltransferase; Amino acids; Bile; Bile acids; Bile Acids and Salts - metabolism; Cell culture; Cell differentiation; Cell lines; Chemical and Drug Induced Liver Injury - metabolism; Cholestasis - metabolism; Cytochrome P450; Dexamethasone; Dimethyl sulfoxide; Enzymes; Hepatocytes; Hepatocytes - metabolism; Hepatoma; Hepatotoxicity; Homeostasis; Humanities and Social Sciences; Humans; Liver; Liver - metabolism; Membrane Transport Proteins - metabolism; multidisciplinary; Organic Anion Transporters, Sodium-Dependent - genetics; Organic Anion Transporters, Sodium-Dependent - metabolism; Science; Science (multidisciplinary); Symporters - metabolism; Taurocholic Acid - metabolism
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-022-18174-z

Hepatic cell lines serve as economical and reproducible alternatives for primary human hepatocytes. However, the utility of hepatic cell lines to examine bile acid homeostasis and cholestatic toxicity is limited due to abnormal expression and function of bile acid-metabolizing enzymes, transporters, and the absence of canalicular formation. We discovered that culturing HuH-7 human hepatoma cells with dexamethasone (DEX) and 0.5% dimethyl sulfoxide (DMSO) for two weeks, with Matrigel overlay after one week, resulted in a shorter and improved differentiation process. These culture conditions increased the expression and function of the major bile acid uptake and efflux transporters, sodium taurocholate co-transporting polypeptide (NTCP) and the bile salt export pump (BSEP), respectively, in two-week cultures of HuH-7 cells. This in vitro model was further characterized for expression and function of bile acid-metabolizing enzymes, transporters, and cellular bile acids. Differentiated HuH-7 cells displayed a marked shift in bile acid composition and induction of cytochrome P450 (CYP) 7A1, CYP8B1, CYP3A4, and bile acid-CoA: amino acid N-acyltransferase (BAAT) mRNAs compared to control. Inhibition of taurocholate uptake and excretion after a 24-h treatment with prototypical cholestatic drugs suggests that differentiated HuH-7 cells are a suitable model to examine cholestatic hepatotoxicity.