Construction of cell sheet-based 3D tissues with designed cell orientation using anisotropic cell sheets

In this study, the cell sheet-based technology was able to control cell orientation in 3D engineered tissue construct. Using a micropatterned thermoresponsive surface, human cells such as fibroblasts and myoblasts were aligned on the surface, and manipulated as a single cell sheet by reducing the culture temperature to 20°C. Consequently, their anisotropic cell sheets can be layered using gelatin gel to produce 3D tissue constructs with the desired anisotropy. For example, two fibroblast sheets layered perpendicularly showed three-dimensionally different cell orientation as designed. By induce differentiation of myoblasts forming an anisotropic cell sheet into myotubes, a myotube construct with a single orientation was possible to be created. Since the combined use of the anisotropic cell sheet and cell sheet manipulation technique allows us to create complex tissue that requires the three-dimensional control of their anisotropies, we believe that it has a potential to be one of the next-generation tissue engineering technology.