High-Throughput and Efficient Fabrication of Engineered Skeletal Muscle Tissue via Streamlined 3D Multimaterial Bioprinting

Abstract

Skeletal muscle diseases have led to an increasing need for a range of physiologically relevant organ-on-a-chip and three-dimensional (3D) culture platforms owing to their clinical significance and ongoing efforts to develop effective therapeutics. However, conventional fabrication approaches still face limitations—such as low throughput, complex fabrication steps, and prolonged production times—making them difficult to apply to efficient drug discovery and development for muscle-related diseases. Here, we present a high-throughput 3D skeletal muscle tissue platform fabricated via a streamlined 3D cell-printing process integrated with a commercially available multi-well plate. Combined with automated 3D printing, the 96-well format enables efficient, rapid, and reproducible fabrication, while significantly reducing processing time and ensuring high tissue uniformity across all wells. To maximize production efficiency, optimized anchor structures and an amphiphilic coating were employed to enhance tissue stability and alignment. In addition, systematic optimization of tissue geometry and 3D bioprinter-based fabrication parameters enabled consistent production of functional and mature 3D skeletal muscle tissues. Overall, this platform integrates 3D bioprinting, optimized anchor design, and controlled tissue geometry to enable a rapid, high-throughput, automation-compatible, and error-minimizing fabrication process. These results demonstrate that the proposed platform can serve as a useful tool for diverse biomedical applications, including the development of effective therapeutics for muscle-related diseases.