PROBLEM TO BE SOLVED: To provide a method for isolating sheet-like cell culture simply and highly efficiently which is used for repairing injured tissues or the like.SOLUTION: A method for producing isolated sheet-like cell culture comprises: freezing the sheet-like cell culture formed by culturing cells seeded on a culture substrate for a period within 24 hours in a condition of adhering to the culture substrate； thawing the freezed sheet-like cell culture by immersing into a medium at 30°C-40°C； and isolating the sheet-like cell culture from the culture substrate. Further, the method for producing the sheet-like cell culture comprises adding a cryopreservation agent to the culture medium, and/or replacing the culture medium by a cryopreservation liquid.
