One-step fabrication of 3D-aligned human skeletal muscle tissue and measurement of contractile force for preclinical drug testing

Human muscle tissue models are critical to understanding the complex physiology of skeletal muscle in studies of drug discovery, development, and toxicity profiling in the human body. However, due to the challenges in in vitro maturation of human muscle cells, few research groups developing their own tissue engineering techniques have succeeded in producing contractile human muscle tissues. Moreover, a more sophisticated method is necessary to measure contractile forces generated by the muscle tissues for preclinical studies in muscle physiology and drug discovery. Although a few research groups have established their own tissue model systems that measure contractile force, they require multi-step fabrication processes to produce human muscle tissues sufficiently functional to be able to measure the contractile forces. To improve the usability of our tissue model system, this study focused on simplifying the tissue engineering approach to produce a practical muscle tissue model. In this study, muscle satellite cells were simply mixed with a combination of fibrinogen, thrombin, and Matrigel before gel formation. The presence of muscle satellite cells induces gel compaction and spontaneously induces unidirectional stretching of the gel, resulting in the muscle satellite cells being aligned threedimensionally with the direction of stretching. Furthermore, this gel environment promotes the maturation of the human muscle progenitor cells into aligned myofibers, also provides the tissue with an elastic platform for muscle contraction, and allows the attachment of the muscle tissue to a device for measurement of contractile force. Therefore, this one-step tissue fabrication allowed us to produce 3D-aligned human muscle tissues and this tissue model is ready to use for the measurement of contractile forces. In fact, the muscle contractions created by electrical and chemical stimulation were quantitatively determined using our measurement system. In addition, the impact of some representative drugs on this muscle tissue were able to be monitored in real-time throughout the changes in contractile forces. In conclusion, our tissue model system, produced by a simple fabrication method, can be used for preclinical in vitro studies in muscle physiology and drug discovery.