Engineered In Vitro Models of Human Skeletal Muscle for Drug Testing, Developmental Biology, and Physiology Studies

Skeletal muscle is the most abundant tissue in the lean human body and is responsible for all motor functions such as locomotion, speech, and respiration. During development, myogenic progenitor cells fuse to form multinucleated muscle fibers that are innervated by the motor neurons of the spinal cord to form functional motor units. Various genetic and acquired diseases of motor neuron, neuromuscular junction (NMJ), and skeletal muscle affect motor unit function with significant effects on patients’ quality of life. Recent advancements in the field of tissue engineering now enable precise control of microenvironment and biochemical cues that emulate physiological conditions and have led to the successful formation of functional human skeletal muscle and neuromuscular tissues in vitro. In 2013 when I initiated my Ph.D. studies, the field had yet to establish parameters to produce engineered human skeletal muscle tissue in a dish capable of responding to electrical and biochemical stimuli. Further, human neuromuscular co-culture models to study NMJ transmission and diseases impacting this critical structure were replete. Since 2013, my work and that of other researchers has overcome barriers to forming functional of human-cell derived skeletal muscle and neuromuscular tissues in vitro. The work in this thesis leveraged, extended, and then applied prior advancements in tissue engineering and engineering techniques, in general, to improve our understanding of skeletal muscle, the neuromuscular junction formation and the maturation of each, in vitro. An important goal of my studies was to encourage widespread adoption of methods I developed by researchers and industry. As such, the tools and technologies were engineered to be simple and integrate into current standard operating procedures and the experimental studies were focused on uncovering biological inquiries that were new and/or were not possible to study with other currently available culture or animal models.Ph.D