Tissue Engineering Strategies for Improved Muscle Regeneration

• James Martin, Medical Diagnostics, University of Delaware

• Brian Kwee, Biomedical Engineering, University of Delaware

Myoblasts are muscle progenitor stem cells that are capable of differentiating and fusing into multinucleated functional units of muscle tissue, known as myotubes. The quality and consistency of lab grown myotubes are heavily dependent on various conditions during cell culture. In my studies, we examined the impact of three of these factors on myogenic differentiation: the number of times myoblasts have been passaged prior to differentiation, the concentration of horse serum used to stimulate differentiation, and the seeding density of myoblasts within the hydrogel scaffolds used to make de novo tissue. These variables are especially relevant to ongoing efforts in tissue engineering, as they are central to all experiments using myoblasts. By making efforts to optimize these parameters, the quality of the myotubes grown in the lab can hopefully be improved. Further increases to the quality of lab grown muscle tissues can not only act to improve the accuracy of research on muscle tissues, but can also lead to the ability to repair heavily damaged muscles with lab grown ones. Moving forward, there are many other factors to investigate that could improve the quality of myotubes formed. These factors include the use of small molecules and antibodies for blocking of specific surface proteins, cell sorting high quality populations of myoblasts, and CRISPR-Cas9 gene editing to modulate key genes involved in myoblast fusion. The ultimate goal is to establish reliable protocols for producing high quality, functional muscle tissue in vitro to advance disease modeling and muscle regeneration strategies.