3D Hepatotoxicity Assay

Detect hepatotoxicity of novel therapeutics with enhanced in vivo relevance using either Cyprotex’s 3D HepaRGTM spheroids and 3D Human Liver Microtissue combined with high content screening (HCS).

Cyprotex delivers consistent, high quality data with the flexibility to adapt protocols based on specific customer requirements.

• Drug induced hepatotoxicity is a leading cause of attrition during drug development. In vitro three-dimensional (3D) cell cultures allow better recapitulation of the complex in vivo microenvironment than traditional 2D monolayer models.
• 3D models also permit long term compound exposures allowing a closer replication of clinical dosing strategies.
• Glutathione depletion, reactive oxygen species (ROS) formation, mitochondrial disruption and cellular ATP depletion are key mechanisms involved in drug induced hepatotoxicity.
• Confocal HCS allows the simultaneous detection of each cell health parameter within a 3D spheroid in combination with a measure of cellular ATP content.

All reference compound toxicities were correctly predicted by the HepaRG^TM spheroid chronic exposure model using the 3D liver toxicity assay with a 5x C_max cut off (table 1). Bosentan and tamoxifen are categorised as false negative responders with ATP alone (table 1) highlighting the enhanced sensitivity of a combined assay. Troglitazone response is one example of the improved assay sensitivity with HCS (ATP MEC 25 µM; DNA MEC 1.69 µM) (figure 2a), while fialuridine highlights the need for long term exposures in vitro (14 day MEC 1.41 µM; 16 hr MEC 451 µM) (figure 2b).

The combination of an in vitro 3D model that better recapitulates the in vivo cellular physiology of hepatic tissue with a multiparametric HCS and cytotoxicity assay presents a viable screening strategy for the accurate in vivo relevant detection of novel therapeutics that cause drug induced liver injury early in drug development.

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