An ordered cellulose film scaffold, termed a nematic ordered cellulose (NOC) template, had unique surface properties and successfully induced the establishment of a three-dimensional (3D), hierarchical structure of epidermal cells by cell attachment and subsequent culture. Initially, the scaffold surface properties were characterized through contact angle measurements and atomic force microscopy to evaluate appropriate hydrophobicity and orientation of molecular chains for 3D culture. The template surfaces exhibited higher hydrophobicity, in the range of 70-75°, than usual cellulose films and appeared suitable for surface cell adhesion. In fact, epidermal cells successfully attached and proliferated favorably on the NOC templates, similar to development in normal culture flasks. Furthermore, the NOC film, as a semipermeable template, was also employed to allow 3D proliferation of epidermal cell layers in the perpendicular direction. The template proved to be suitable as a 3D cell culture device, resulting in the proposal that the construction processes of these 3D cell layers followed the basic concept of skin formation.