Methods of cell culture for adoptive cell therapy

A method for culturing and then concentrating cells in a gas permeable cell culture and cell recovery device comprising:

• adding an initial volume of medium and cells into the gas permeable cell culture and cell recovery device;
• said gas permeable cell culture and cell recovery device comprising a cell culture compartment bounded by a growth surface and a wall that is perpendicular to said growth surface, said growth surface comprised of liquid impermeable gas permeable material, the bottom of said gas permeable material in contact with a growth surface support that holds the gas permeable material in a horizontal position during culture and allows ambient gas to contact the underside of the gas permeable material by random motion, said wall is more than 2.0 cm in height;
• wherein the initial volume of media resides at an initial medium height determined by the distance from the lowermost medium location to the uppermost medium location;
• and wherein the initial medium height is beyond 2.0 cm, and the ratio of said initial volume of media to the growth surface area is beyond 2.0 mL/cm2 and the medium is not perfused;
• allowing the cells to gravitate to the growth surface and allowing the cells to reside in a state of static culture without medium replenishment for a period of time with said gas permeable cell culture and cell recovery device in a location with ambient gas suitable for cell culture such that the cells expand;
• concentrating the cells by inserting a medium removal conduit into the gas permeable cell culture and cell recovery device and removing medium to obtain a cell recovery medium volume and a cell recovery medium height, wherein the cell recovery medium height is 0.2 cm or more, and the cell recovery volume to growth surface area ratio is 0.2 ml/cm2 or more.