title: 3D organotypic cultures of human HepaRG cells: a tool for in vitro toxicity studies.
abstract: Drug-induced human hepatotoxicity is difficult to predict using the current in vitro systems. In this study, long-term 3D organotypic cultures of the human hepatoma HepaRG cell line were prepared using a high-throughput hanging drop method. The organotypic cultures were maintained for 3 weeks and assessed for (1) liver specific functions, including phase I enzyme and transporter activities, (2) expression of liver-specific proteins, and (3) responses to three drugs (acetaminophen, troglitazone, and rosiglitazone). Our results show that the organotypic cultures maintain high liver-specific functionality during 3 weeks of culture. The immunohistochemistry analyses illustrate that the organotypic cultures express liver-specific markers such as albumin, CYP3A4, CYP2E1, and MRP-2 throughout the cultivation period. Accordingly, the production rates of albumin and glucose, as well as CYP2E1 activity, were significantly higher in the 3D versus the 2D cultures. Toxicity studies show that the organotypic cultures are more sensitive to acetaminophen- and rosiglitazone-induced toxicity but less sensitive to troglitazone-induced toxicity than the 2D cultures. Furthermore, the EC50 value (2.7mM) for acetaminophen on the 3D cultures was similar to in vivo toxicity. In summary, the results from our study suggest that the 3D organotypic HepaRG culture is a promising in vitro tool for more accurate assessment of acute and also possibly for chronic drug-induced hepatotoxicity.
authors: Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, 171 77 Stockholm, Sweden.
authors: Patrina
authors: P
authors: Gunness
authors: Daniel
authors: D
authors: Mueller
authors: Valery
authors: V
authors: Shevchenko
authors: Elmar
authors: E
authors: Heinzle
authors: Magnus
authors: M
authors: Ingelman-Sundberg
authors: Fozia
authors: F
authors: Noor
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000378:metabolism; D000082:Acetaminophen / Q000633:toxicity; D000418:Albumins / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D002839:Chromans / Q000633:toxicity; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000378:metabolism; D000072467:Cytochrome P450 Family 2; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D008099:Liver / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D000089762:Multidrug Resistance-Associated Protein 2; D000077154:Rosiglitazone; D018874:Spheroids, Cellular / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D045162:Thiazolidinediones / Q000633:toxicity; D023382:Toxicity Tests, Acute / Q000379:methods*; D000077288:Troglitazone; D014508:Urea / Q000378:metabolism; D000097202:ATP-Binding Cassette Sub-Family B Member 4
chemical_list: D018435:ATP Binding Cassette Transporter, Subfamily B; D000082:Acetaminophen; D000418:Albumins; D002839:Chromans; D003577:Cytochrome P-450 Enzyme System; D000072467:Cytochrome P450 Family 2; D000089762:Multidrug Resistance-Associated Protein 2; D000077154:Rosiglitazone; D045162:Thiazolidinediones; D000077288:Troglitazone; D014508:Urea; D000097202:ATP-Binding Cassette Sub-Family B Member 4; C000715087:ABCC2 protein, human; C110332:CYP2F1 protein, human