Enhanced Oxygen Cell Culture Platforms

• US 20080009027A1
• Filed: 07/09/2007
• Published: 01/10/2008
• Est. Priority Date: 07/07/2006
• Status: Active Grant

• Current Assignee: University of Miami
• Original Assignee: University of Miami
• Inventors: Inverardi, Luca, Fraker, Christopher, Camillo, Ricordi, Dominguez-Bendala, Juan
• Granted Patent: US 8,551,770 B2
• Time in Patent Office: Days
• US Class Current: 435/029
• CPC Class Codes: C12M 23/04 Flat or tray type, drawers C12M23/10, C12M23/12, C12M23/16 take precedence
• 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

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 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 membrane barrier;

support members extending from the bottom, side or top of the tissue culture well to elevate the tissue culture well; and

,

a tray or culture flask comprising the tissue culture well.

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

and a gas permeable, liquid-impermeable membrane barrier wherein said barrier defines a continuous bottom surface the tissue culture device.

3. The cell and tissue culture apparatus of claim 1, wherein the membrane barrier comprises at least one material selected from 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, hydrogels and silicone.

4. The cell and tissue culture apparatus of claim 3, wherein the membrane barrier comprises a perfluorinated hydrocarbon and silicone composition.

5. The cell and tissue culture apparatus of claim 3, wherein the perfluorinated hydrocarbon comprises fluoroheptanes, fluorocycloheptanes, fluoromethylcycloheptanes, fluorohexanes, fluorocyclohexanes, fluoropentanes, fluorocyclopentanes, fluoromethylcyclopentanes, fluorodimethylcyclopentanes, fluoromethylcyclobutanes, fluorodimethylcyclobutanes, fluorotrimethylcyclobutanes, fluorobutanes, fluorocyclobutanse, fluoropropanes, fluoroethers, fluoropolyethers, fluorotributylamines, fluorotriethylamines, perfluorohexanes, perfluoropentanes, perfluorobutanes, perfluoropropanes, or sulfur hexafluoride.

6. The cell and tissue culture apparatus of claim 4, wherein the perfluorinated hydrocarbon and silicone composition has a ratio of between about 0.001% v/v perfluorinated hydrocarbon per ml of silicone up to 80% v/v perfluorinated hydrocarbon per ml of silicone.

7. The cell and tissue culture apparatus of claim 4, wherein the perfluorinated hydrocarbon and silicone composition has a ratio of about 1% to 20% v/v perfluorinated hydrocarbon per ml of silicone.

8. The cell and tissue culture apparatus of claim 4, wherein the membrane barrier comprises pores of at least about 0.001 μ

m up to 1 mm in size.

9. The cell and tissue culture apparatus of claim 1, wherein the tissue culture device 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, hydrogels and silicone.

10. 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, hydrogels and silicone.

11. The cell and tissue culture apparatus of claim 1, wherein the membrane barrier is at least about 0.01 μ

M thick up to about 1 mm thick.

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

13. The cell and tissue culture apparatus of claim 12, wherein the tray comprises a 6-wells, 12-wells, 24-wells, 96-wells or 384 wells.

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

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

(a) 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;

(b) introducing the suspension into an instrument for injecting the suspension into a tissue culture well of the apparatus; and

,

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

24. The method of claim 23, wherein the cells to be cultured are anchorage-dependent cells.

25. The method of claim 23, wherein the cells to be cultured are anchorage-independent cells.

26. The method of claim 23 wherein the cells are stem cells.

15. A cell and tissue culture apparatus in the shape of the bag comprising a perfluorinated hydrocarbon and silicone composition of at least about 0.01 μ

M thick up to about 1 mm thick.

16. The cell and tissue culture apparatus of claim 15, wherein the bag has at least one opening.

17. The cell and tissue culture apparatus of claim 15, wherein the bag comprises one or more chambers.

18. A composition comprising a highly permeable silicone matrix impregnated with a perfluorocarbon.

19. The composition of claim 18, wherein the perfluorocarbon comprises fluoroheptanes, fluorocycloheptanes, fluoromethylcycloheptanes, fluorohexanes, fluorocyclohexanes, fluoropentanes, fluorocyclopentanes, fluoromethylcyclopentanes, fluorodimethylcyclopentanes, fluoromethylcyclobutanes, fluorodimethylcyclobutanes, fluorotrimethylcyclobutanes, fluorobutanes, fluorocyclobutanse, fluoropropanes, fluoroethers, fluoropolyethers, fluorotributylamines, fluorotriethylamines, perfluorohexanes, perfluoropentanes, perfluorobutanes, perfluoropropanes, or sulfur hexafluoride.

20. The composition of claim 18, wherein the perfluorocarbon and silicone composition has a ratio of between about 0.001% v/v perfluorocarbon per ml of silicone up to 80% v/v perfluorocarbon per ml of silicone.

21. The composition of claim 18, wherein the perfluorocarbon and silicone composition has a ratio of about 20% v/v perfluorocarbon per ml of silicone.

22. The composition of claim 18, wherein the perfluorocarbon and silicone composition has a ratio of about 10% v/v perfluorocarbon per ml of silicone.

27. A method to determine the effect of enhanced oxygen delivery and availability to a cell culture, said method comprising culturing cells in an apparatus comprising:

a tissue culture device containing a membrane barrier;

support members extending from the bottom, side or top of the tissue culture well to elevate the tissue culture well; and

,

a tray or culture flask comprising the tissue culture well.

28. The method of claim 27, wherein the apparatus comprises an upper and lower opening;

and a gas permeable, liquid impermeable membrane barrier wherein said barrier defines a continuous bottom surface the tissue culture device.

29. The method of claim 27, wherein the membrane barrier comprises at least one material selected from 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, hydrogels and silicone.

30. The method of claim 27, wherein the membrane barrier comprises a perfluorinated hydrocarbon and silicone composition.

31. The method of claim 27, wherein the perfluorinated hydrocarbon comprises fluoroheptanes, fluorocycloheptanes, fluoromethylcycloheptanes, fluorohexanes, fluorocyclohexanes, fluoropentanes, fluorocyclopentanes, fluoromethylcyclopentanes, fluorodimethylcyclopentanes, fluoromethylcyclobutanes, fluorodimethylcyclobutanes, fluorotrimethylcyclobutanes, fluorobutanes, fluorocyclobutanse, fluoropropanes, fluoroethers, fluoropolyethers, fluorotributylamines, fluorotriethylamines, perfluorohexanes, perfluoropentanes, perfluorobutanes, perfluoropropanes, or sulfur hexafluoride.

34. The apparatus of claim 31, wherein the membrane barrier comprises at least one material selected from ceramics, polymers, woven substrates, non-woven substrates, polyamide, polyester, polyurethane, fluorocarbon polymers, polyethylene, polypropylene, perfluorinated hydrocarbon, polyvinyl alcohol, hydrogels and silicone.

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

a culture device containing a membrane barrier; and

,

a container enclosing the culture device.

33. The apparatus of claim 32, wherein the culture device comprises an upper and lower opening;

and a gas permeable membrane barrier wherein said barrier defines a continuous surface of at least one side of the culture device.

35. The apparatus of claim 32, wherein the membrane barrier comprises a perfluorinated hydrocarbon and silicone composition.

36. The apparatus of claim 32, wherein the perfluorinated hydrocarbon comprises fluoroheptanes, fluorocycloheptanes, fluoromethylcycloheptanes, fluorohexanes, fluorocyclohexanes, fluoropentanes, fluorocyclopentanes, fluoromethylcyclopentanes, fluorodimethylcyclopentanes, fluoromethylcyclobutanes, fluorodimethylcyclobutanes, fluorotrimethylcyclobutanes, fluorobutanes, fluorocyclobutanse, fluoropropanes, fluoroethers, fluoropolyethers, fluorotributylamines, fluorotriethylamines, perfluorohexanes, perfluoropentanes, perfluorobutanes, perfluoropropanes, or sulfur hexafluoride.

37. The apparatus of claim 36, wherein the perfluorinated hydrocarbon and silicone composition has a ratio of between about 0.001% v/v perfluorinated hydrocarbon per ml of silicone up to 80% v/v perfluorinated hydrocarbon per ml of silicone.

38. The apparatus of claim 35, wherein the perfluorinated hydrocarbon and silicone composition has a ratio of about 1% to 20% v/v perfluorinated hydrocarbon per ml of silicone.

39. The apparatus of claim 35, wherein the membrane barrier comprises pores of at least about 0.001 μ

m up to 1 mm in size.

40. The apparatus of claim 35, wherein the culture device and container are 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, hydrogels and silicone.

41. The apparatus of claim 36, wherein the membrane barrier is at least about 0.01 μ

M thick up to about 1 mm thick.