Extracellular matrix-immobilized nanotopographical substrates for enhanced myogenic differentiation

Extracellular matrix (ECM) components such as fibronectin (FN) and laminin (LMN) play prominent roles in controlling cellular behaviors. Many attempts have been made to explore cellular behaviors on combinatorial ECM arrays in high-throughput systems. However these studies were limited to physical adsorption of ECM, which does not guarantee a lasting effect of ECM in vitro. Here, we demonstrate ECM immobilization on polyurethane acrylate (PUA) substrate fabricated in 24 well-plate platforms to effectively differentiate C2C12. Our study demonstrate that co-immobilization of FN and LMN was found to enhance myogenic differentiation of C2C12 cells compared to single immobilization of either FN or LMN alone. Furthermore, utilizing nano-imprint lithography technique, 300 nm and 5 µm line-patterned substrates were fabricated on 24-well plates. FN and LMN co-immobilized substrates with line-patterns additionally provided the directionality for mimicking musculoskeletal structure and enhanced the myogenic differentiation.