Human iPSC-derived Hepatocytes and Cardiomyocytes for Drug Toxicity Testing
Poster Mar 30, 2015
AnnandRR; Vardaro R; Hamilton B; Akakira R; Tamura K; Yoshida S; Lin YC; Toyoda D; Kogami H; Okuda Y; Watanabe T; Inamura M
Pharmaceutical candidate compounds require an extended period of time and high development costs before reaching the market. However, for various reasons, a high number of the candidate compounds are eliminated in the development process. The candidate compound elimination is frequently based on hepatotoxicity and cardiotoxicity. Although human primary cells are widely used for drug toxicity testing, they have posed issues such as lot-to-lot variation. Moreover, it is difficult to perform long-term tests with the same donor when using human primary cells due to the limited supply.
Human iPSC-derived hepatocytes ReproHepato™ cells were cultivated on 96-well plates, and the cells were exposed to five representative toxic compounds: acetaminophen, amiodarone, cyclophosphamide, diclofenac, and flutamide. Using conventional assays to measure ATP and LDH, we observed dose-dependent toxicity, and each compound showed IC50 values similar to the cytotoxicity observed using primary human hepatocytes (PHHs). Furthermore, we also observed toxicity using high content analysis, which simultaneously measures cell number, reduction in glutathione level, active oxygen, and mitochondrial membrane potential. These results show that ReproHepato can be used as an alternative to PHHs for routine cytotoxicity testing.
We cultivated Human iPSC-derived cardiomyocytes ReproCardio 2™ cells on 96-well plates, and exposed the cells to five representative toxic compounds: aspirin, verapamil, isoproterenol, E-4031 and flecainide. Upon the addition of chemical compounds, the prolongation or reduction of the field potential duration was observed in a Multi-Electrode Array (MEA) experiment, with small lot-to-lot variation. These results demonstrate that ReproCardio 2 is a suitable model system for in vitro cardiotoxicity screening for QT prolongation and arrhythmia potential.
In summary, human iPSC-derived hepatocytes and cardiomyocytes can be used for drug toxicity testing, as alternative sources of human primary cells.