Highly efficient gas permeable devices and methods for culturing cells

Abstract

This invention relates to methods and devices that improve cell culture efficiency. They include the use of gas permeable culture compartments that reduce the use of space while maintaining uniform culture conditions, and are more suitable for automated liquid handling. They include the integration of gas permeable materials into the traditional multiple shelf format to resolve the problem of non-uniform culture conditions. They include culture devices that use surfaces comprised of gas permeable, plasma charged silicone and can integrate traditional attachment surfaces, such as those comprised of traditional tissue culture treated polystyrene. They include culture devices that integrate gas permeable, liquid permeable membranes. A variety of benefits accrue, including more optimal culture conditions during scale up and more efficient use of inventory space, incubator space, and disposal space. Furthermore, labor and contamination risk are reduced.

1 . A method for culturing cells in a static cell growth apparatus, said apparatus comprising:
 a liquid impermeable housing, the inside of which is able to contain cells and medium and the outside of which is in contact with ambient gas; and   said housing forming a plurality of gas permeable shelves, each having an inside surface and an outside surface; and   the inside surface of each shelf having an opposing surface located a distance away and defining a culture space; and   the culture spaces are located one above the other when the shelves are in a horizontal position; and   a manifold that connects the culture spaces; and   projections that make contact with the outside surface of each shelf while leaving a portion of the outside surface in contact with ambient gas;   the method comprising:   adding cells and a volume of liquid medium to the apparatus;   orienting the apparatus into a first position such that the shelves are one above the other;   allowing cells to settle upon multiple shelves while the apparatus is oriented in the first position;   adding enough liquid medium to prevent a unique gas-liquid interface from forming directly above at least one shelf when the device is oriented in the first position;   having at least a portion of the liquid medium in contact with at least a portion of at least one gas permeable material;   placing the cell culture device in a cell culture location that includes ambient gas at a composition suitable for cell culture, said ambient gas making contact with said gas permeable material; and   
       not perfusing the apparatus when the apparatus is in said cell culture location. 
     
     
         2 . The method of  claim 1  wherein said cell culture device is further adapted so that a portion of said liquid medium resides in a portion of said cell culture device other than directly above said scaffolds in the first position. 
     
     
         3 . The method of  claim 1  wherein a unique gas-liquid interface does not form directly above more than one scaffold when the device is oriented in the first position. 
     
     
         4 . The method of  claim 1  wherein a unique gas-liquid interface does not form directly above any scaffold when the device is oriented in the first position. 
     
     
         5 . The method of  claim 1 , whereby said housing is capable of being completely filled with media for optimal cell-nutrient exchange. 
     
     
         6 . The apparatus of  claim 1 , whereby a uniformity of conditions for cellular growth includes a determined media volume per unit surface area. 
     
     
         7 . A method for culturing cells in a static cell growth apparatus, said apparatus comprising:
 a liquid impermeable housing, the inside of which is able to contain cells and medium and the outside of which is in contact with ambient gas; and   said housing defining a plurality of gas permeable shelves, each having an inside surface and an outside surface; and   the inside surface of each shelf having an opposing surface located a distance away and defining a culture space; and   the culture spaces are located one above the other when the shelves are in a horizontal position; and   a manifold that connects the culture spaces; and   projections that make contact with the outside surface of each shelf while leaving a portion of the outside surface in contact with ambient gas;   the method comprising:   adding cells and a volume of liquid medium to the apparatus;   orienting the apparatus into a first position such that the shelves are one above the other;   allowing cells to settle upon multiple shelves while the apparatus is oriented in the first position;   adding enough liquid medium to prevent a unique gas-liquid interface from forming directly above at least one shelf when the device is oriented in the first position;   having at least a portion of the liquid medium in contact with at least a portion of at least one gas permeable material;   placing the cell culture device in a cell culture location that includes ambient gas at a composition suitable for cell culture, said ambient gas making contact with said gas permeable material; and   
       not perfusing the apparatus when the apparatus is in said cell culture location. 
     
     
         8 . The method of  claim 7  wherein said cell culture device is further adapted so that a portion of said liquid medium resides in a portion of said cell culture device other than directly above said scaffolds in the first position. 
     
     
         9 . The method of  claim 7  wherein a unique gas-liquid interface does not form directly above more than one scaffold when the device is oriented in the first position. 
     
     
         10 . The method of  claim 7  wherein a unique gas-liquid interface does not form directly above any scaffold when the device is oriented in the first position. 
     
     
         11 . The method of  claim 7 , whereby said housing is capable of being completely filled with media for optimal cell-nutrient exchange. 
     
     
         12 . The apparatus of  claim 7 , whereby a uniformity of conditions for cellular growth includes a determined media volume per unit surface area.