High-content analysis of in vitro hepatocyte injury induced by various hepatotoxicants
- Author:
Nga T T THAM
1
;
So Ryeon HWANG
;
Ji Hyun BANG
;
Hee YI
;
Young Il PARK
;
Seok Jin KANG
;
Hwan Goo KANG
;
Yong Sang KIM
;
Hyun Ok KU
Author Information
- Publication Type:Original Article
- Keywords: High-content analysis; Multi-parameter cell-based assay; HepG2 cell; Human primary hepatocyte; Hepatotoxicity
- MeSH: Animals; Calcium; Cytosol; Fluorescence; Hep G2 Cells; Hepatocytes; Humans; In Vitro Techniques; Lipid Peroxidation; Liver; Membrane Potential, Mitochondrial
- From:Journal of Veterinary Science 2019;20(1):34-42
- CountryRepublic of Korea
- Language:English
- Abstract: In vitro prediction of hepatotoxicity can enhance the performance of non-clinical animal testing for identifying chemical hazards. In this study, we assessed high-content analysis (HCA) using multi-parameter cell-based assays as an in vitro hepatotoxicity testing model using various hepatotoxicants and human hepatocytes such as HepG2 cells and human primary hepatocytes (hPHs). Both hepatocyte types were exposed separately to multiple doses of ten hepatotoxicants associated with liver injury whose mechanisms of action have been described. HCA data were obtained using fluorescence probes for nuclear size (Hoechst), mitochondrial membrane potential (TMRM), cytosolic free calcium (Fluo-4AM), and lipid peroxidation (BODIPY). Cellular alterations were observed in response to all hepatotoxicants tested. The most sensitive parameter was TMRM, with high sensitivity at a low dose, next was BODIPY, followed by Fluo-4AM. HCA data from HepG2 cells and hPHs were generally concordant, although some inconsistencies were noted. Both hepatocyte types showed mild or severe mitochondrial impairment and lipid peroxidation in response to several hepatotoxicants. The results demonstrate that the application of HCA to in vitro hepatotoxicity testing enables more efficient hazard identification, and further, they suggest that certain parameters could serve as sensitive endpoints for predicting the hepatotoxic potential of chemical compounds.
