Multiparametric High-Content Assays to Measure Cell Health and Oxidative Damage as a Model for Drug-Induced Liver...

Area of Science:

• Biochemistry
• Cell Biology
• Toxicology

Background:

• Drug-induced liver injury (DILI) is a major challenge in drug development, leading to drug non-approval and market withdrawal.
• Current in vitro toxicity screening relies heavily on hepatic cell lines but lacks comprehensive health markers.
• There is a need for advanced assays to predict DILI and elucidate toxicity mechanisms.

Purpose of the Study:

• To develop and validate novel live cell-based assays for assessing hepatotoxicity.
• To measure diverse cell health parameters indicative of liver injury using HepG2 cells.
• To enhance the predictive value of in vitro assays for preclinical drug safety.

Main Methods:

• Developed three live cell-based assays using HepG2 cells.
• Assay 1: Luciferase-based measurement of cellular ATP levels.
• Assays 2 & 3: Multiparametric high-content screening of cell count, mitochondrial function, nuclear morphology, vacuolar density, reactive oxygen species, and glutathione levels.

Main Results:

• Successfully implemented ATP measurement assay for cell viability.
• High-content assays quantified multiple cell health parameters simultaneously.
• Demonstrated the capability to assess a broad range of toxicity indicators in HepG2 cells.

Conclusions:

• The described assays provide a more comprehensive assessment of hepatotoxicity than traditional methods.
• These advanced in vitro assays can improve the prediction of drug-induced liver injury.
• The multiparametric approach aids in understanding the mechanisms of drug toxicity during preclinical development.