Development of In Vitro Drug-Induced Cardiotoxicity Assay by Using Three-Dimensional Cardiac Tissues Derived from Human Induced Pluripotent Stem Cells

• Published in: Tissue engineering. Part C, Methods Vol. 24; no. 1; pp. 56 - 67
• Main Authors: Takeda, Maki, Miyagawa, Shigeru, Fukushima, Satsuki, Saito, Atsuhiro, Ito, Emiko, Harada, Akima, Matsuura, Ryohei, Iseoka, Hiroko, Sougawa, Nagako, Mochizuki-Oda, Noriko, Matsusaki, Michiya, Akashi, Mitsuru, Sawa, Yoshiki
• Format: Journal Article
• Language: English
• Published: United States Mary Ann Liebert, Inc 01.01.2018
• Subjects: Antibiotics, Antineoplastic - adverse effects; Artificial tissues; Assaying; Cardiomyocytes; Cardiotoxicity; Cardiotoxins - adverse effects; Cell Survival; Cells, Cultured; Collagen (type I); Cytotoxicity; Doxorubicin; Doxorubicin - adverse effects; Drug discovery; Drug Evaluation, Preclinical - methods; Drug screening; Drugs; Extracellular matrix; Fibroblasts; Fibronectin; Gelatin; Growth factors; Heart; Heart diseases; Heart surgery; HERG protein; Humans; Induced Pluripotent Stem Cells - cytology; Isoproterenol; L-Lactate dehydrogenase; Lactic acid; Laminin; Medicine; Muscle contraction; Muscle Contraction - drug effects; Myocytes; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - metabolism; Myocytes, Cardiac - pathology; Pluripotency; Potassium; Potassium channels (voltage-gated); Proteins; Stem cells; Studies; Three dimensional imaging; Tissues; Toxicity; University graduates; 3D cardiac tissue; drug screening; cardiotoxicity
• ISSN: 1937-3384; 1937-3392
• DOI: 10.1089/ten.tec.2017.0247

An in vitro drug-induced cardiotoxicity assay is a critical step in drug discovery for clinical use. The use of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is promising for this purpose. However, single hiPSC-CMs are limited in their ability to mimic native cardiac tissue structurally and functionally, and the generation of artificial cardiac tissue using hiPSC-CMs is an ongoing challenging. We therefore developed a new method of constructing three-dimensional (3D) artificial tissues in a short time by coating extracellular matrix (ECM) components on cell surfaces. We hypothesized that 3D cardiac tissues derived from hiPSC-CMs (3D-hiPSC-CT) could be used for an in vitro drug-induced cardiotoxicity assay. 3D-hiPSC-CT were generated by fibronectin and gelatin nanofilm coated single hiPSC-CMs. Histologically, 3D-hiPSC-CT exhibited a sarcomere structure in the myocytes and ECM proteins, such as fibronectin, collagen type I/III, and laminin. The administration of cytotoxic doxorubicin at 5.0 μM induced the release of lactate dehydrogenase, while that at 2.0 μM reduced the cell viability. E-4031, human ether-a-go-go related gene (hERG)-type potassium channel blocker, and isoproterenol induced significant changes both in the Ca transient parameters and contractile parameters in a dose-dependent manner. The 3D-hiPSC-CT exhibited doxorubicin-sensitive cytotoxicity and hERG channel blocker/isoproterenol-sensitive electrical activity in vitro , indicating its usefulness for drug-induced cardiotoxicity assays or drug screening systems for drug discovery.