Centrifugation chamber with gas permeable membrane layers for cell cultivation
The invention is directed to a centrifuge chamber comprising a cylinder having a base plate and a cover plate, a rotational axis with at least one port for input and/or output of liquids, at least one port for input of gases and at least one layer for cell culturing, wherein the layer for cell culturing comprises a gas-permeable membrane on which cells can be cultured and wherein the at least one port for input and/or output of gases is connected to the gas-permeable membrane.
1. A centrifuge chamber having a rotational axis, an inner circumference and an outer wall, comprising:
a base plate;
a cover plate;
at least one liquid port for liquids;
at least one gas port for gases; and
at least one layer for cell culturing, wherein each layer for cell culturing comprises a gas-permeable membrane on which cells are cultured, and a rigid support that supports the gas-permeable membrane, and wherein the both the gas-permeable membrane and the rigid support each have a through hole structure formed therein, wherein the through hole structure in the membrane fits over the through hole structure in the rigid support, thereby defining their relative orientation, and wherein the membrane further has spacers that allow the flow of gas from the through hole structures to a volume between the membrane and the rigid support, and wherein the base plate is coupled to a gas port and to at least one gas permeable membrane, so that the gas port is in gaseous communication with the at least one gas permeable membrane, wherein each layer has at least two through hole structures on the membrane and rigid support, wherein the at least two through hole structures are about 180 degrees apart on the surface of the membrane or rigid structure, and wherein one of the two through hole structures is sealed on each layer, defining a serpentine gas path for the gas through the centrifuge chamber.
2. The centrifuge chamber of claim 1 , wherein the through hole structure in the rigid support has a narrower dimension on one side than an obverse side, such that the through hole structure one layer can accept the through hole structure on an adjacent layer in an interlocking, stacked structure.
3. The centrifuge chamber according to claim 1 , wherein the support structure is provided with channels.
4. The centrifuge chamber of claim 1 , wherein the gas-permeable membrane has a gas-tight seal at its inner and outer diameter against the rigid support, forming an enclosed volume between the gas-permeable membrane and the rigid support.
5. The centrifuge chamber according to claim 1 , wherein the gas-permeable membrane is coupled to the rigid layer and the base plate in a gas tight manner.
6. The centrifuge chamber according to claim 1 , wherein the gas-permeable membrane and the support structure have an optical transparency suitable for optical microscopy.
7. The centrifuge chamber according to claim 1 , wherein a surface of the gas-permeable membrane is functionalized for cell culturing.
8. The centrifuge chamber according to claim 1 , wherein the at least one layer for cell culturing has a diameter at least 5% smaller than a diameter of the outer wall of the chamber.
9. The centrifuge chamber according to claim 1 , wherein the at least one liquid port is connected to an opening in at least one of the base plate and cover plate and the opening is provided with at least one deflector.
10. The centrifuge chamber according to claim 9 , wherein the at least one deflector has a width at its base of at most 1/10 of the inner circumference of the chamber.
11. The centrifuge chamber according to claim 9 , wherein the at least one deflector is substantially parallel with the outer wall of the chamber and has a width of 5 to 50 times the width of the opening at which it is located.
12. The centrifuge chamber according to claim 11 , wherein a first deflector is located between at least one opening and the rotational axis of the cylinder and a second deflector is located between at least one opening and the outer wall of the chamber.
13. A centrifuge chamber having a rotational axis, an inner circumference and an outer wall, comprising:
a base plate;
a cover plate;
at least one liquid port for liquids;
at least one gas port for gases; and
at least one layer for cell culturing, wherein each layer for cell culturing comprises a gas-permeable membrane on which cells are cultured, and a rigid support that supports the gas-permeable membrane, and wherein the both the gas-permeable membrane and the rigid support each have a through hole structure formed therein, wherein the through hole structure in the membrane fits over the through hole structure in the rigid support, thereby defining their relative orientation, and wherein the membrane further has spacers that allow the flow of gas from the through hole structures to a volume between the membrane and the rigid support, and wherein the base plate is coupled to a gas port and to at least one gas permeable membrane, so that the gas port is in gaseous communication with the at least one gas permeable membrane, wherein a plurality of layers for cell culturing is disposed on top of one another in an interlocked stack, and wherein each layer for cell culturing comprises at least one opening interconnecting the layers in the stack and wherein the openings of two adjacent layers are located opposite each other and on a line intersecting the rotational axis, and wherein gas circulates through the stack in a serpentine fashion.
14. The centrifuge chamber of claim 13 , wherein a first and last layer of the stack comprise openings covered with sterile filters across a direction of gas flow.
15. The centrifuge chamber according to claim 13 , wherein a first layer of the stack is connected to at least one gas port.