Disclosed are a method for producing a frozen pharmaceutical composition prepared from cultured human mesenchymal stem cells and a pharmaceutical composition containing human mesenchymal stem cells. This method is a method for producing a frozen pharmaceutical composition containing human mesenchymal stem cells, which comprises the following steps in this order: (a) adding trypsin to human mesenchymal stem cells in a culture vessel to detach the cells from the surface of the culture vessel； (b) adding a bicarbonate Ringer's solution containing human serum albumin to the detached cells to terminate the reaction with trypsin, and washing the cells with the bicarbonate Ringer's solution containing human serum albumin； (c) suspending the cells in a bicarbonate Ringer's solution containing human serum albumin and dimethyl sulfoxide； (d) putting the resulting suspension in a container which allows freezing of what is contained therein, and sealing the container； and (e) freezing the suspension put in the container.
