Human iPSC-engineered cardiac tissue platform faithfully models important cardiac physiology

• Published in: American journal of physiology. Heart and circulatory physiology Vol. 320; no. 4; pp. H1670 - H1686
• Main Authors: de Lange, Willem J, Farrell, Emily T, Kreitzer, Caroline R, Jacobs, Derek R, Lang, Di, Glukhov, Alexey V, Ralphe, J Carter
• Format: Journal Article
• Language: English
• Published: United States American Physiological Society 01.04.2021
• Series: Deconstructing Organs: Single-Cell Analyses, Decellularized Organs, Organoids, and Organ-on-a-Chip Models
• Subjects: Adrenergic beta-Agonists - pharmacology; Animal models; Calcium; Calcium (intracellular); Calcium - metabolism; Calcium Signaling - drug effects; Cardiomyocytes; Cell culture; Cell Differentiation; Cell Line; Contraction; Electroconvulsive therapy; Embryos; Energy resources; Fibrin; Fibroblasts; Handling; Heart; Humans; Induced Pluripotent Stem Cells - drug effects; Induced Pluripotent Stem Cells - metabolism; Induced Pluripotent Stem Cells - ultrastructure; Isoforms; Kinetics; Maturation; Muscle contraction; Myocardial Contraction; Myocardium; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - metabolism; Myocytes, Cardiac - ultrastructure; Pharmacology; Phenotype; Physiology; Pluripotency; Stem cell transplantation; Stem cells; Stimuli; Substrates; Tissue Engineering; Tissues; Transcription; Ventricle; t-tubules; maturation; human iPS cells; contractility; engineered cardiac tissue
• ISSN: 0363-6135; 1522-1539
• DOI: 10.1152/AJPHEART.00941.2020

Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CM) may provide an important bridge between animal models and the intact human myocardium. Fulfilling this potential is hampered by their relative immaturity, leading to poor physiological responsiveness. hiPSC-CMs grown in traditional two-dimensional (2D) culture lack a t-tubular system, have only rudimentary intracellular calcium-handling systems, express predominantly embryonic sarcomeric protein isoforms, and preferentially use glucose as an energy substrate. Culturing hiPSC-CM in a variety of three-dimensional (3D) environments and the addition of nutritional, pharmacological, and electromechanical stimuli have proven, to various degrees, to be beneficial for maturation. We present a detailed assessment of a novel model in which hiPSC-CMs and hiPSC-derived cardiac fibroblasts are cocultured in a 3D fibrin matrix to form engineered cardiac tissue constructs (hiPSC-ECTs). The hiPSC-ECTs are responsive to physiological stimuli, including stretch, frequency, and β-adrenergic stimulation, develop a t-tubular system, and demonstrate calcium-handling and contractile kinetics that compare favorably with ventricular human myocardium. Furthermore, transcript levels of various genes involved in calcium-handling and contraction are increased. These markers of maturation become more robust over a relatively short period of time in culture (6 wk vs. 2 wk in hiPSC-ECTs). A comparison of the hiPSC-ECT molecular and performance variables with those of human cardiac tissue and other available engineered tissue platforms is provided to aid selection of the most appropriate platform for the research question at hand. Important and noteworthy aspects of this human cardiac model system are its reliance on "off-the-shelf" equipment, ability to provide detailed physiological performance data, and the ability to achieve a relatively mature cardiac physiology without additional nutritional, pharmacological, and electromechanical stimuli that may elicit unintended effects on function. This study seeks to provide an in-depth assessment of contractile performance of human iPSC-derived cardiomyocytes cultured together with fibroblasts in a 3-dimensional-engineered tissue and compares performance both over time as cells mature, and with corresponding measures found in the literature using alternative 3D culture configurations. The suitability of 3D-engineered human cardiac tissues to model cardiac function is emphasized, and data provided to assist in the selection of the most appropriate configuration based on the target application.