Coupling 3D Printing and Novel Replica Molding for In House Fabrication of Skeletal Muscle Tissue Engineering Devices

The transition from 2D to 3D engineered tissue cultures is changing the way biologists can perform in vitro functional studies. However, there has been a paucity in the establishment of methods required for the generation of microdevices and cost‐effective scaling up. To date, approaches including multistep photolithography, milling and 3D printing have been used that involve specialized and expensive equipment or time‐consuming steps with variable success. Here, a fabrication pipeline is presented based on affordable off‐the‐shelf 3D printers and novel replica molding strategies for rapid and easy in‐house production of hundreds of 3D culture devices per day, with customizable size and geometry. This pipeline is applied to generate tissue engineered skeletal muscles in vitro using human induced pluripotent stem cell‐derived myogenic progenitors. These production methods can be employed in any standard biomedical laboratory. A pipeline based on 3D printing and replica molding to fabricate polydimethylsiloxane (PDMS)‐based devices for skeletal muscle tissue engineering is presented. The pipeline involves three demolding methods: simple peeling, dissolving the printed mold, and creating a highly elastic intermediate mold. Devices with different size, levels of complexity, and throughput can be produced, while maintaining comparable biological characteristics in the engineered tissues.