Amsterdam University Medical Center 1 article published in JoVE Bioengineering Generation, High-Throughput Screening, and Biobanking of Human-Induced Pluripotent Stem Cell-Derived Cardiac Spheroids Renee G. C. Maas1, Tess Beekink*1, Nino Chirico*1, Christian J. B. Snijders Blok1, Inge Dokter1, Vasco Sampaio-Pinto1, Alain van Mil1, Pieter A. Doevendans1, Jan W. Buikema2, Joost P. G. Sluijter*1, Francesca Stillitano*1 1Utrecht Regenerative Medicine Center, Circulatory Health Laboratory, University Utrecht, Department of Cardiology, University Medical Center Utrecht, 2Amsterdam Cardiovascular Sciences, Department of Physiology, Amsterdam University Medical Center Presented here is a set of protocols for the generation and cryopreservation of cardiac spheroids (CSs) from human-induced pluripotent stem cell-derived cardiomyocytes cultured in a high-throughput, multidimensional format. This three-dimensional model functions as a robust platform for disease modeling, high-throughput screenings, and maintains its functionality after cryopreservation.