Maintenance of high quality rat precision cut liver slices during culture to study hepatotoxic responses: Acetaminophen as a model compound

• Published in: Toxicology in vitro Vol. 42; pp. 200 - 213
• Main Authors: Granitzny, Anne, Knebel, Jan, Schaudien, Dirk, Braun, Armin, Steinberg, Pablo, Dasenbrock, Clemens, Hansen, Tanja
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.08.2017; Elsevier Science Ltd
• Subjects: Acetaminophen; Acetaminophen - toxicity; Adenosine Triphosphate - metabolism; Albumin; Albumins - metabolism; Analgesics; Animals; Aspartate transaminase; ATP; Chemical and Drug Induced Liver Injury - metabolism; Chemical and Drug Induced Liver Injury - pathology; Correlation analysis; Culture; Culture conditions; Culture setup; Dehydrogenase; Glutamate dehydrogenase; Hepatotoxicity; L-Lactate dehydrogenase; Lactate dehydrogenase; Lactic acid; Liver; Liver - drug effects; Liver - metabolism; Liver - pathology; Liver diseases; Male; Models, Biological; Organ Culture Techniques; Precision cut liver slices; Proteins; Rats, Wistar; Toxicity; Toxicology; Transaminase; Urea; Urea - metabolism; Viability; Cmax; SER; AST; DILI; PCLiS; Culture conditions; GLDH; Precision cut liver slices; BSA; LDH; SD; Culture setup; Acetaminophen; EC50; KH; Hepatotoxicity; NL
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2017.05.001

Precision cut liver slices (PCLiS) represent a promising tool in reflecting hepatotoxic responses. However, the culture of PCLiS varies considerably between laboratories, which can affect the performance of the liver slices and thus the experimental outcome. In this study, we describe an easily accessible culture method, which ensures optimal slice viability and functionality, in order to set the basis for reproducible and comparable PCLiS studies. The quality of the incubated rat PCLiS was assessed during a 24h culture period using ten readouts, which covered viability (lactate dehydrogenase-, aspartate transaminase- and glutamate dehydrogenase-leakage, ATP content) and functionality parameters (urea, albumin production) as well as histomorphology and other descriptive characteristics (protein content, wet weight, slice thickness). The present culture method resulted in high quality liver slices for 24h. Finally, PCLiS were exposed to increasing concentrations of acetaminophen to assess the suitability of the model for the detection of hepatotoxic responses. Six out of ten readouts revealed a toxic effect and showed an excellent mutual correlation. ATP, albumin and histomorphology measurements were identified as the most sensitive readouts. In conclusion, our results indicate that rat PCLiS are a valuable liver model for hepatotoxicity studies, particularly if they are cultured under optimal standardized conditions. •A high overall viability and functionality can be maintained in rat PCLiS for 24h.•APAP-treatment of rat PCLiS showed that DILI can be reflected comprehensively.•High quality rat PCLiS lead to highly reproducible and comparable results.