A method of producing a cell culture chip, in which a first adhesive is film-formed on a first substrate having transparency such that a first adhesive layer and a recessed portion on which one or more cell culture flow paths are formed face each other, and a second adhesive is film-formed on the first substrate such that a second adhesive layer and a flat portion around the recessed portion face each other, thereby obtaining a cell culture flow path substrate, the first adhesive containing a polyester-based resin having a glass transition temperature of 5° C. or higher, and the second adhesive containing a polyester-based resin having a glass transition temperature of lower than 5° C.