Abstract: Human-induced pluripotent stem cell (hiPSC)-derived cardiac patches show promise for myocardial infarction treatment but are limited by insufficient thickness. Previously, we developed a fibrous scaffold that supported the formation of well-organized cardiac tissue constructs. In this study, based on the above technology, we developed a three-dimensional multilayer fibrous scaffold with dynamic perfusion, allowing the single-step seeding of approximately 20 million hiPSC-derived cardiomyocytes (CMs) to form 1 mm thick, viable tissue. This multilayer tissue exhibited enhanced contractility, increased cytokine secretion, and significantly improved functional recovery with reduced fibrosis in a myocardial infarction model. These findings indicate the need for precise evaluation of hiPSC-CM dosage in clinical therapy development, as higher cell doses significantly enhanced therapeutic outcomes.
