A polystyrene scaffold based human three-dimensional liver model for drug discovery

Introduction Cell culture monolayer (2D) models have widely been used as test systems in drug discovery. With respect to the biological function organ characteristics are limited and less pronounced compared to the in vivo situation. Cells grown in a three-dimensional (3D) environment demonstrated physiological characteristics that are closer to In vivo. Here we report on a 3D liver model as surrogate to follow adverse effects induced by drugs. Objectives The objective was to demonstrate that the established 3D liver model runs liver toxicity by Acetaminophen (AAP) similarly compared to the in vivo situation. Methods HepaRG cells were seeded into polystyrene scaffold (PS) and cultured for one week. To differentiate, cells were treated with 1% DMSO for 28 days followed by AAP treatment. To prevent the AAP-induced hepatotoxicity, the 3D liver model was pretreated with 5 mM N-Acetylcysteine (NAC). Following parameters were determined: reactive oxygene species (ROS) (dihydrorhodamine 123), mitochondrial membrane potential (JC-1 assay), Glutathione (GSH assay), and cytotoxicity (LDH assay). Results Liver parameters were significantly higher in the 3D liver model vs. 2D. AAP increased ROS and NAC counteracts, decreased significantly mitochondrial membrane potential, and metabolic activity and Glutathione level up to 500 μM after 24 h incubation time. Conclusion The 3D liver model is regarding the AAP-induced hepatotoxicity close to the in vivo situation. Therefore, it is a predictive new system for in vitro test applications to study liver function in drug metabolism, hepatotoxicity and may contribute to reduce animal experimentation.

Keywords:

3D cell culture , Liver model , Polystyrene , Scaffold , Drug discovery