SOT 62nd Annual Meeting and ToxExpo
Venue: Nashville Music City Center
in downtown Nashville, Tennesse
Join us at the SOT 62nd Annual Meeting and ToxExpo
Visit us at booth #543 and meet Ronald Knox (Nanion Senior Business Executive) and our friends from innoVitro at the SOT meeting and learn about AtlaZ hepatotox-screening on FCDI hepatocytes.
Information about the poster presentation
Poster number: #4286
Title: Development of a high-throughput system for advanced hepatotoxicity screening
Presenter (s): Ronald Knox in collaboration with Fujifilm Cellular Dynamics
Drug-Induced Liver Injury (DILI) still remains the major cause of acute liver failure. The toxicants act through different mechanisms and with different kinetics, hence there is a need to assess the risk of drug-induced hepatotoxicity, allowing a phenotypic and kinetic understanding of these toxicological responses. Ideally, such an approach should, at same time, meet high throughput demands of the medical community, to efficiently identify hepatic toxicity and to allow a fast development and launch of new disease therapies.
Our aim was to develop an easy-to-use system, serving high throughput hepatotoxicity screening demands. iCell Hepatocytes 2.0 were treated with reference compounds with known hepatotoxic effects and were measured on our technology. Furthermore, responses were compared to existing cell viability and toxicity assay results on the same cells.
We adopted 96-well electric cell-substrate impedance sensing technology (CardioExcyte 96) and implemented multiple data acquisition frequencies ranging from 0.1 kHz – 100 kHz. Additionally, we scaled up the throughput from 1 x 96 wells to the newly developed 6 x 96-wells system (AtlaZ).
96-well culture plates with integrated gold-film electrodes reveal information on confluency, cell-cell interaction and conductivity of adherent cells and thereby provide a measure of toxicity. Data acquisition at different recording frequencies allows detection of diverse cell properties and greatly enhances the information content obtained in each well. For example, high frequency impedance is highly sensitive to differences in cell-confluency, making it useful for measuring cell growth or proliferation rates and cytotoxicity, whereas low frequency impedance detects changes in the space under or between cells and therefore enables barrier function and cell adhesion quantification.
Our data revealed that the cultured hepatocytes were metabolically active and functional when the impedance-based, label-free and non-invasive assay was used. Acute and chronic (dose-dependent) liver toxic effects have been tested for 5 relevant compounds: diclofenac, APAP, Troglitazone, Chlorpromazine and Aflatoxin B1. To obtain 2D monolayers in 96-well plates, cells were seeded with density of 300k cells/cm2, and recordings were performed during the time period of 21 days.
Data showed comparable results to other cell viability and tox assays, such as CellTiter-Glo, proving impedance as a reliable but non-invasive tool. We demonstrated dose and time dependent effects for all 5 compounds on iCell Hepatocytes 2.0. Importantly, the acute and chronic effects of Aflatoxin B1 were observed in standard 2D conditions, indicating that the hepatocytes were metabolically active and functional when using this impedance-based, non-invasive assay.
An upscaling of the recording device to allow for 6 x 96 measurements in a simultaneous or independent manner was performed.
The use of impedance and electrical impedance spectroscopy emerges as a valuable tool for advanced hepatotoxicity assessment. The newly developed AtlaZ system elevates preclinical risk assessment to a modern level through the scalable throughput and ease-of-use, but also through the potential to access multiple kinetic and phenotypic information from in vitro 2D cell cultures, exemplarily shown here with iCell Hepatocytes 2.0.