Disclosed are a method for manufacturing a cell sheet, comprising culturing cells on a fibrin-coated surface of a substrate until the cells reach confluency, continuing the cultivation of the cells for a sufficient time period to cause the degradation of fibrin at the bottom of the cells, and detaching the cultured cells from the substrate surface in a sheet-like form to give a cell sheet; a substrate for cell sheet preparation, a surface of which is coated with fibrin; and a composition for use in coating with fibrin a surface of a substrate for cell sheet preparation, the composition comprising fibrinogen and thrombin. The invention enables to cell sheets to be manufactured by a simple manipulation using a substrate coated with a commercial, commonly available material of which safety has been confirmed.