Liver Cell Culture on ITO-SAMs and Metabolite Analysis using NMR for Drug Toxicity Applications
Description
Despite the emergence of three-dimensional (3D) cell culture systems, two dimensional (2D) cultures remain prevalent due to their cost-effectiveness, reproducibility, and ease of analysis. This study aims to develop a novel self-assembled monolayer (SAM)-based cell culture platform for growth of hepatocytes and to explore advanced metabolomic analysis techniques for enhanced understanding of hepatotoxicity. Indium-Tin Oxide (ITO) functionalized with 3-aminopropyl (triethoxysilane) (APTES) is hypothesized to serve as an optimal scaffold for drug toxicity screening and as a model for Micro Engineered Organs (MOE). Surface characterization was performed using Atomic Force Microscopy (AFM) and X-ray Photoelectron Spectroscopy (XPS). HepG2 liver cells were cultured on ITO modified with APTES. Cell proliferation and cytotoxicity were assessed using MTT and Live/Dead assays. The scaffold of ITO modified with APTES provides an optimal SAM surface for HepG2 cell adhesion and proliferation as evidenced by confocal microscopy images. Preliminary metabolomic analysis through Nuclear Magnetic Resonance (NMR) spectroscopy offers detailed insights into intracellular responses and metabolites normally undetectable by regular CMP (comprehensive metabolomic pathway) tests. This study lays the groundwork for the development of more physiologically relevant in vitro hepatocyte models, potentially enhancing the accuracy of drug toxicity screening.