Enhanced oxygen cell culture platforms

• US 9,175,254 B2
• Filed: 10/07/2013
• Issued: 11/03/2015
• Est. Priority Date: 07/07/2006
• Status: Active Grant

• Current Assignee: University of Miami
• Original Assignee: University of Miami
• Inventors: Dominguez-Bendala, Juan, Ricordi, Camillo, Inverardi, Luca, Fraker, Christopher A.
• Primary Examiner(s): Beisner, William H
• Assistant Examiner(s): HENKEL, DANIELLE B
• Application Number:
  US14/047,218
• Publication Number:
  US 20140099717A1
• Time in Patent Office: 757 Days
• US Class Current: 1/1
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/20 Material Coatings immunocoatings C12M25/00
• CPC Class Codes: C12M 23/24 Gas permeable parts
• CPC Class Codes: C12M 25/04 in combination with well or multiwell plates, i.e. culture inserts
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis

Abstract

Provided is a cell culture apparatus for culturing cells, that provides enhanced oxygen delivery and supply to cells without the need for stirring or sparging. Oxygen diffusion occurs on both sides of the culture vessel, top and bottom. A gas-permeable membrane that includes perfluorocarbons or fluorocarbon derivatives (e.g., fluorinated silane, partially fluorinated silane) in its composition allows for the rapid, enhanced and uniform transfer of oxygen between the environment of cells or tissues contained in the cell culture container apparatus and the atmosphere of the incubator in which the cell culture apparatus is incubated.

Claims

1. A cell and tissue culture apparatus comprising:

a tissue culture device containing a gas permeable, liquid impermeable membrane barrier, the gas permeable, liquid impermeable membrane comprising a silicone composition impregnated, mixed or combined with a fluorocarbon derivative;

support members extending from the bottom, side or top of the tissue culture device to elevate the tissue culture device to allow air flow; and

, a tray or culture flask comprising the tissue culture device, wherein the liquid impermeable membrane barrier defines a continuous surface of the tissue culture device and allows oxygenation of and minimization of hypoxia in cells resting thereupon.

2. The cell and tissue culture apparatus of claim 1, wherein the fluorocarbon derivative is fluorosilane.

3. The cell and tissue culture apparatus of claim 2, wherein the fluorosilane is triethoxyfluorosilane.

4. The cell and tissue culture apparatus of claim 1, wherein the tissue culture apparatus comprises an upper and lower opening;

and wherein the barrier defines a continuous bottom surface of the tissue culture device.

5. The cell and tissue culture apparatus of claim 1, wherein the liquid impermeable membrane barrier further comprises at least one material selected from ceramics, polymers, woven substrates, non-woven substrates, polyamide, polyester, polyurethane, fluorocarbon polymers, polyethylene, polypropylene, polyvinyl alcohol, polystyrene, vinyl, plastics, metals, alloys, minerals, non-metallic minerals, wood, fibers, cloth, glass, and hydrogels.

6. The cell and tissue culture apparatus of claim 1, wherein the fluorocarbon derivative and silicone composition has a ratio of between about 0.001% v/v fluorocarbon derivative per ml of composition up to 80% v/v fluorocarbon derivative per ml of silicone composition.

7. The cell and tissue culture apparatus of claim 6, wherein the fluorocarbon derivative and silicone composition has a ratio of about 10% to 20% v/v fluorocarbon derivative per ml of silicone composition.

8. The cell and tissue culture apparatus of claim 1, wherein the tray is made from at least one material comprising ceramics, polymers, woven substrates, non-woven substrates, polyamide, polyester, polyurethane, fluorocarbon polymers, polyethylene, polypropylene, perfluorinated hydrocarbon, polyvinyl alcohol, polystyrene, vinyl, plastics, metals, alloys, minerals, non-metallic minerals, wood, fibers, cloth, glass, hydro gels and silicone.

9. The cell and tissue culture apparatus of claim 1, wherein the tray comprises a plurality of tissue culture devices.

10. The cell and tissue culture apparatus of claim 1, wherein the tray comprises a lid which allows for gaseous exchange.

11. The cell and tissue culture apparatus of claim 1, wherein the oxygenation and minimization of hypoxia increases proliferation of the cells.

12. The cell and tissue culture apparatus of claim 1, wherein the oxygenation and minimization of hypoxia increases differentiation of the cells.

13. The cell and tissue culture apparatus of claim 1, wherein the cells are selected from the group consisting of:

neural stem cells, beta cells and hepatocytes.

14. The cell and tissue culture apparatus of claim 1, wherein the fluorocarbon derivative is homogenously dispersed throughout the silicone composition.

15. A method of growing cells or tissue explants in an enhanced oxygen delivery tissue culture apparatus, the method comprising:

suspending the cells or tissue explants to be cultured in the apparatus according to claim 1, in an appropriate amount of tissue culture medium to form a suspension; and

incubating the cell culture apparatus, containing the suspension of medium and cells, in a cell culture incubator.

16. The method of claim 15, wherein the cells to be cultured are anchorage-dependent cells.

17. The method of claim 15, wherein the cells to be cultured are anchorage-independent cells.

18. The method of claim 15, wherein the cells are stem cells.

19. An apparatus for the transportation of cells, tissues and organs comprising:

a culture device containing a gas permeable, liquid impermeable membrane barrier to allow air flow, the gas permeable, liquid impermeable membrane comprising a silicone composition impregnated, mixed or combined with a fluorocarbon derivative,

wherein the liquid impermeable membrane barrier defines a continuous surface of at least one side of the culture device and allows oxygenation of and minimization of hypoxia in cells resting thereupon; and

,

a container enclosing the culture device.