Fabrication of 3D Cardiac Microtissue Arrays using Human iPSC-Derived Cardiomyocytes, Cardiac Fibroblasts, and Endothelial Cells

Generation of human cardiomyocytes (CMs), cardiac fibroblasts (CFs), and endothelial cells (ECs) from induced pluripotent stem (iPSCs) has provided a unique opportunity to study the complex interplay among different cardiovascular cell types that drives tissue development disease. In area models, several sophisticated three-dimensional (3D) approaches use cell-derived (iPSC-CMs) mimic physiological relevance native environment with combination extracellular matrices crosslinkers. However, these systems are fabricate without microfabrication expertise require weeks self-assemble. Most importantly, many lack vascular make up over 60% nonmyocytes in heart. Here we describe derivation all three iPSCs microtissues. This facile replica molding technique allows microtissue culture standard multi-well plates for weeks. The platform user-defined control sizes based on initial seeding density requires less than 3 days self-assembly achieve observable contractions. Furthermore, microtissues can be easily digested while maintaining high viability single-cell interrogation flow cytometry RNA sequencing (scRNA-seq). We envision this vitro model will help accelerate validation studies drug discovery disease modeling.