Two-dimensional differential adherence and alignment of C2C12 myoblasts

Surface microstructure is a critical parameter for scaffolds used in skeletal muscle tissue engineering. We have developed micromachined surfaces using matrix assisted pulsed laser evaporation-direct write (MAPLE-DW) that demonstrate differential adherence of C2C12 myoblast cells. The 60-400 μm diameter channels were micromachined onto 2% agarose surfaces using an ArF excimer laser and lined with Matrigel® basement membrane matrix solution. Suspensions containing C2C12 myoblast cells were then placed on the surface of these micromachined channels. The C2C12 myoblast cells aligned themselves parallel to the 60-150 μm channels. Live/dead assays over 72 h demonstrated that cell number, cell size, and number of nuclei per cell increased within these channels. In addition, some of the myoblasts fused and differentiated into multinucleated myotubes. These results provide the basis for the development of direct-write scaffolds for skeletal muscle tissue engineering.