Abstract: It is an object of the present invention to provide a method for producing frozen mesenchymal cells maintaining a viability and an adhesion rate similar to those of cells obtained through a step of culture, without undergoing a step of culturing cells before cryopreservation step. The method comprises dissociating cells by treating a tissue with proteolytic enzymes and cryopreserving the cells. The present invention provides a method for producing frozen mesenchymal cells maintaining a viability and an adhesion rate similar to those of culturing cells without a step of culturings cells before cryopreserving.