Design and Validation of a 3D-Printed Muscle Stimulation Chamber for In Vitro Muscle Tissue Culture

Abstract

Skeletal muscle contraction is essential not only for generating force but also for triggering vital biological and biochemical responses. Studying these processes in situ is challenging due to difficulties in achieving precise control and measurement of mechanochemical regulation in the complex in vivo microenvironment. This research aimed at developing and validating a 3D-printed electrical pulse stimulation chamber specifically designed for in vitro skeletal muscle culture, utilizing the C2C12 murine myoblast cell line. The primary objective was to create a low-cost, modular platform that allows precise control over stimulation parameters such as voltage, frequency, and pulse duration to investigate the mechanisms of muscle contraction. Previous studies using existing electrical stimulation systems lacked scalability, compatibility with incubators, and cost-effectiveness. Therefore, the first aim of this research was to design a chamber that allows customizable control over these parameters. The second aim involved evaluating the cytocompatibility of the materials used in the 3D printing process and the established electrical stimulation circuitry for their use in cell culture. The final aim was to assess the chamber’s effectiveness in promoting key biological processes in muscle cells, particularly differentiation and the production of the myokine interleukin-6 (IL-6). Validation of the electrical stimulation system demonstrated its stability and reliability, with consistent electrical output over a 24-hour period. Significant increases in IL-6 production in electrically stimulated myotubes confirmed the system's capacity to elicit physiological responses. This 3D-printed platform presents a tool for further studies into muscle physiology and has the potential to advance muscle cell culture applications for better understanding muscle contraction, regeneration, and repair.