Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) as a new source of cardiac cells are expected to find use as tools in high-throughput screening for drug candidates and cardiotoxicity validation without the need for experimentation on animals. In recent years, it has been reported that drug screening using three-dimensional (3D) tissue is better than conventional 2D culture. Various methods have been developed for mass culture of hiPSC-CMs, and embryoid body (EB) formation is necessary in the majority of differentiation methods as this is reported to promote the differentiation of hiPSCs. However, these operations result in increased processing, cost and loss of hiPSCs. Here, we show alternative methods for differentiation to hiPSC-CMs from <100 mu m hiPSC-clumps without EB formation and report on a 3D-tissue fabrication using hiPSC-CMs. The 3D cardiac tissue constructed by a layer-by-layer (LbL) cell coating technique (LbL-3D Heart) showed synchronous powerful beating. We conclude that this method enables cost-effective, reproducible and scalable hiPSC-CM production with high activity for tissue engineering, drug screening and regenerative medicine. (C) 2020, The Society for Biotechnology, Japan. All rights reserved.