Compartmentalized multiple well tissue culture plate

A static self contained cell culture device consisting essentially of a container having:

• (a) a plurality of cell culture compartments (40) each including a first gas permeable film (120) disposed below and an upper sheet (20) selectively permeable to compounds of less than a particular size, means separating said first gas permeable film (120) from said upper sheet (20) to allow a culture medium (50) to reside between said upper sheet (20) and said first gas permeable film (120);
• b) means defining a basal medium compartment (30) above said cell culture compartments (40) to allow a basal medium (60) to reside in contact with said upper sheet (20);
• c) an access port (70) to each of said cell culture compartments (40);
• d) an access port (90) to said basal medium compartment;
• e) a gas film support (130) below and in partial contact with said first gas permeable film (120) suspension cells can distribute across said gas permeable film (120) and gas transfer into and out of said cell culture compartment (40) is not substantially impaired;
• f) a second gas permeable film (230) disposed in a horizontal position below said first gas permeable film (120);
• g) separating means between said first gas permeable film (120) and said second gas permeable film (230) to form a variable oxygen control compartment (220) adapted to contain fluid;
• and
• h) an access port (250) to said variable oxygen control compartment (220) whereby liquid can be added or removed to control the rate of gas transport.

A method of culturing cells comprising the steps:

• a) forming a plurality of cell culture compartments (40) each of said cell culture compartments (40) comprising of a gas permeable film (120) disposed below an upper sheet (20) selectively permeable to compounds of less than a particular size;
• b) forming a gas film support (130) disposed below said gas permeable film (120) and adapted to hold said gas permeable film (120) in a substantially horizontal position while allowing gas to contact the underside of said gas permeable film (120);
• c) placing a basal medium (60) in a self contained basal medium compartment (30) above at least one cell culture compartment (40), said basal medium (60) communicating with cell culture medium (50) by way of said upper sheet (20) maintaining said basal medium in a substantially quiescent state;
• d) placing cells and a cell culture medium (50) in at least one of said cell culture compartments (40), said cell culture medium (50) in communication with basal medium (60) by way of said upper sheet (20);
• e) maintaining said cells at a selected temperature;
• and
• f) allowing gas exchange through said gas permeable film (120), whereby cells are allowed to proliferate upon the upper surface of said gas permeable film (120).

A method of culturing cells comprising the steps:

• a) forming a plurality of cell culture compartment (40), each of said cell culture compartments (40) comprising a gas permeable, liquid permeable film (120) disposed below an upper sheet (20) selectively permeable to compounds of less than a particular size;
• b) placing a basal medium (60) in at least one compartment above a cell culture compartment (40), said basal medium (60) contacting with said upper sheet (20);
• c) placing the cells and a cell culture medium (50) in at least one of said cell culture compartments (40), said cell culture medium (50) communicating with basal medium (60) by way of said upper sheet (20);
• d) maintaining said cells at an appropriate temperature;
• and
• e) allowing gas to reside below said gas permeable, liquid permeable film (120).

A method of culturing cells comprising the steps;

• a) forming a plurality of cell culture compartment (40), each of said cell culture compartments (40) comprising a first gas permeable film (120) disposed below an upper sheet (20) permeable to compounds of less than a particular size;
• b) placing said cell culture compartment (40) upon a variable oxygen control compartment (220) comprised of a bottom second gas permeable film (230) disposed below an upper gas film support (130), maintaining said first gas permeable film (120) of said cell culture compartment (40) in a substantially horizontal position;
• c) placing a basal medium (60) in a self contained basal medium compartment (30) above at least one of said cell culture compartment (30), said basal medium (60) contacting with said upper sheet (20);
• d) placing cells and a cell culture medium (50) in said cell culture compartment (40), said cell culture medium (50) communicating with basal medium (60) by way of said upper sheet (20);
• e) placing a selected volume of a liquid (260) into said variable oxygen control compartment (220);
• f) maintaining said cells at a predetermined temperature;
• and
• g) allowing oxygen and carbon dioxide to diffuse through said gas permeable film (120) by way of said liquid (260) and said bottom second gas permeable film (230) of said variable oxygen control compartment (220), whereby cells are allowed to proliferate upon the upper surface of said first gas permeable film 120, and
• h) optionally varying the oxygen tension within said cell culture compartment 40 by adding or removing said liquid (260) from said variable oxygen control compartment (40).

A method of culturing cells comprising the steps of:

• a) forming a plurality of cell culture compartment (40), each of said cell culture compartments (40) comprising a gas permeable film (120) disposed below an upper sheet (20) selectively permeable to compounds of less than a particular size;
• b) forming an access port (70) to each of said formed cell culture compartments (40);
• c) maintaining said gas permeable film (120) in a substantially horizontal position;
• d) placing a basal medium (60) in a self contained basal medium compartment (40), said basal medium (60) contacting said upper sheet (20) maintaining said basal medium in a substantially quiescent state;
• e) placing cells and a cell culture medium (50) in at least one of said cell culture compartments (40), said culture medium (50) communicating with said basal medium (60) by way of upper sheet (20);
• f) allowing a pressure equilibrium to be established across said upper sheet (20) of said cell culture compartments (40) by allowing the liquid level in said culture compartment to rise into said access port of said cell culture compartment to counter balance the hydrostatic pressure of said basal medium;
• g) maintaining said cells at a selected temperature;
• and
• h) allowing gas exchange through said gas permeable film (120) whereby cells are allowed to proliferate upon the upper surface of said gas permeable film (120).