A novel and simple method for constructing cell sheets is described in which culture surfaces were modified with RGD peptide-coupled alginate hydrogels using an electrodeposition method. Cells were cultured on the hydrogels to form a contiguous cell sheet-like structure. Then, the hydrogels were dissolved by addition of an EDTA solution and the cell sheets rapidly detached from the surface. This is the first report of the fabrication of cell sheets on RGD peptide-coupled alginate hydrogels using an electrodeposition method. We believe the technique is useful for cell sheet engineering.