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 comprising:
 adding cells and media to said apparatus;   wherein the apparatus comprises only one compartment,   said only one compartment comprises multiple culture spaces; and   said compartment is liquid impermeable, capable of holding medium, and   includes more than one gas permeable shelf for cells to reside upon; and   each shelf has a culture space between it and an opposing surface; and   a manifold connects each culture space, and the culture spaces reside one above the other when the shelves are in a horizontal position; and   said apparatus comprises projections capable of supporting the outside surface of each shelf and said projections are spaced apart to create a space for ambient gas to contact each shelf; and   placing the apparatus in a tissue culture incubator.   
     
     
         2 . The method of  claim 1 , wherein the media and cells completely fill said compartment. 
     
     
         3 . The method of  claim 1 , wherein said opposing surface is gas permeable, liquid impermeable. 
     
     
         4 . The method of  claim 1 , whereby a uniformity of conditions for cellular growth includes a determined media volume per unit surface area. 
     
     
         5 . The method of  claim 1 , wherein said compartment has a substantially rectangular footprint and a substantially uniform height. 
     
     
         6 . A method of culturing cells in cell growth apparatus comprising:
 adding cells and media to said cell growth apparatus,   wherein said apparatus does not have pumps or other equipment to perfuse medium or gas;   said apparatus is comprised of multiple components;   said apparatus comprising a liquid impermeable structure, the inside of which is able to contain cells and medium and the outside of which is in contact with ambient gas; and   more than one surface on the inside of said liquid impermeable structure being in contact with multiple gas permeable shelves each gas permeable shelf having an inside surface and an outside surface; and   the inside surface of each gas permeable shelf having an opposing surface located a distance away, defining a culture space between each gas permeable shelf and its opposing surface;   each culture space having side walls and an opening that allows media and cells to enter and exit the culture space, and   the culture spaces are located one above the other when the gas permeable shelves are in a horizontal position; and   a manifold that connects the openings of each culture space;   projections that are adapted to make contact with the outside surface of each gas permeable shelf when the device is in use, while leaving a portion of the outside surface of each gas permeable shelf in contact with ambient gas; and   culturing cells in said apparatus in an incubator.   
     
     
         7 . The method of  claim 6 , whereby said liquid impermeable structure is completely filled with cells and media for optimal cell-nutrient exchange. 
     
     
         8 . The method of  claim 6 , wherein said opposing surface is gas permeable, liquid impermeable. 
     
     
         9 . The method of  claim 6 , whereby a uniformity of conditions for cellular growth includes a determined media volume per unit surface area. 
     
     
         10 . A method for culturing cells in a cell growth apparatus comprising:
 adding cells and media into a cell growth apparatus that is comprised of a single liquid impermeable compartment that includes multiple culture spaces, each of which includes one gas permeable surface for cells to reside upon and an opposing surface; and   said apparatus includes a manifold that connects each culture space to allow media and cells to go into each culture space;   said apparatus includes a space for gas to make contact with the gas permeable surface of each culture space, and   placing the apparatus in a tissue culture incubator wherein the culture spaces reside one above the other and culturing the cells in the presence of ambient incubator gas and without perfusing gas or liquid through the apparatus at any time when it is in the incubator.   
     
     
         11 . The method of  claim 1 , wherein said media and cells completely fill said compartment. 
     
     
         12 . The method of  claim 1 , wherein said opposing surface is gas permeable, liquid impermeable. 
     
     
         13 . The method of  claim 1 , whereby a uniformity of conditions for cellular growth includes a determined media volume per unit surface area. 
     
     
         14 . The method of  claim 1 , wherein said compartment has a substantially rectangular footprint and a substantially uniform height.