In vitro model for cardiac organoid production

This study presents a novel, cost-effective, high-throughput method to generate cardiac organoids (COs) from induced pluripotent stem cells (iPSCs) using substrates of known stiffness, geometrical confinement, and Wnt signaling modulation. By day 21, COs exhibited key cardiac features, including spontaneous contraction, multicellularity, and chamber-like structures. Characterization was performed using multi-omics (bulk/single-cell RNA-seq, proteomics), immunofluorescence, electrophysiology, and contraction measurements. Environmental stiffness significantly influenced cardiac maturity and ultrastructure. Functional assays confirmed ionic currents, spontaneous action potentials, and pharmacological responsiveness, highlighting the potential of COs for in vitro drug testing and disease modeling.