The three dimensional organoids-based high content imaging model for hepatotoxicity assessment / 药学学报

ABSTRACT

The drug hepatotoxicity assessment method in vitro was established by 3D organoid model of HepaRG cell line in combination with high content imaging analysis. HepaRG cells were differentiated into hepatocyte-like morphology and bile canaliculus-like structures by treatment with hydrocortisone and dimethyl sulfoxide(DMSO), inducing the expressions of drug-metabolizing enzymes, transporters, nuclear receptors and hepatocyte-specific protein albumin(ALB)genes, finally forming the stable organoids with closely resembling liver function in vitro. Through the high content imaging analysis and the specific, multi-targets fluorescent dye, the number of live/dead cells, mitochondrial membrane potential(MMP), intracellular reactive oxygen species(ROS)were analyzed for the drug hepatotoxicity evaluation. The results showed that the organoids evaluation model of HepaRG cells in vitro could be used to assess accurately the difference between hepatotoxicity positive control drugs of amiodarone(AMD), cyclosporin(CSP)and the negative control drug of aspirin(ASP) AMD and CSP concentration-dependently decreased the number of total and live organoid cells. The number of dead organoid cells was increased sharply when the concentration of AMD was more than 50 μmol·L-1, while no significant changes was observed for ASP. AMD and CSP concentration-dependently caused the MMP declined and the ROS increased, with AMD showing a greater degree than CSP and ASP presenting no markedly effect. In conclusion, the organoid evaluation method of HepaRG cells in combination with high content imaging analysis can be used for the drug hepatotoxicity assessment in vitro. It displays the advantages of multi-target, high throughput, intuitive results as well as quantitatively.