Cell culture methods and devices utilizing gas permeable materials

• US 9,441,192 B2
• Filed: 07/27/2015
• Issued: 09/13/2016
• Est. Priority Date: 10/08/2003
• Status: Active Grant

• Current Assignee: Wilson Wolf Manufacturing Corporation
• Original Assignee: Wilson Wolf Manufacturing Corporation
• Inventors: Wilson, John R., Page, Douglas A., Welch, Daniel, Robeck, Alison
• Primary Examiner(s): Beisner, William H
• Application Number:
  US14/810,071
• Publication Number:
  US 20160024465A1
• Time in Patent Office: 414 Days
• US Class Current: 1/1
• CPC Class Codes: C12M 23/08 Flask, bottle or test tube
• CPC Class Codes: C12M 23/24 Gas permeable parts
• CPC Class Codes: C12M 25/06 Plates; Walls; Drawers; Multilayer plates
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12N 2500/02 Atmosphere, e.g. low oxygen conditions
• CPC Class Codes: C12N 5/00 Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor; plant reproduction by tissue culture techniques A01H4/00
• CPC Class Codes: C12N 5/0602 Vertebrate cells

Abstract

Gas permeable devices and methods are disclosed for cell culture, including cell culture devices and methods that contain medium at heights, and certain gas permeable surface area to medium volume ratios. These devices and methods allow improvements in cell culture efficiency and scale up efficiency.

Claims

1. A method of culturing cells comprising:

adding medium and animal cells into a static cell culture device that is not compartmentalized by a semi-permeable membrane, at least a portion of said cell culture device is comprised at least in part of a non porous gas permeable material, ambient gas is in contact with at least a portion of said gas permeable material, and

placing said cell culture device in a cell culture location that includes ambient gas at a composition suitable for animal cell culture, wherein said cell culture device is oriented in a position such that at least a portion of said cells reside upon at least a portion of said gas permeable material, the uppermost location of said medium is elevated beyond 2.0 cm from the lowermost location of said medium, and said device is in a state of static cell culture.

2. The method of claim 1 wherein the uppermost location of said medium is elevated at least 3.2 cm from the lowermost location of said medium.

3. The method of claim 1 wherein the uppermost location of said medium is elevated at least 4.0 cm from the lowermost location of said medium.

4. The method of claim 1 wherein the uppermost location of said medium is elevated at least 5.09 cm from the lowermost location of said medium.

5. The method of claim 1 wherein the uppermost location of said medium is elevated at least 6.0 cm from the lowermost location of said medium.

6. The method of claim 1 wherein the uppermost location of said medium is elevated at least 7.0 cm from the lowermost location of said medium.

7. The method of claim 1 wherein the uppermost location of said medium is elevated at least 8.0 cm from the lowermost location of said medium.

8. A method of culturing cells comprising:

adding medium and animal cells into a static cell culture device that is not compartmentalized by a semi-permeable membrane, the bottom of said device is comprised at least in part of a non porous gas permeable material, and

placing said device in a cell culture location that includes ambient gas at a composition suitable for animal cell culture, wherein said non porous gas permeable material is in a horizontal plane, the uppermost location of said medium is more than 2.0 cm above the lowermost location of said medium, at least a portion of said gas permeable material is in contact with ambient gas, and said device is not subjected to mixing or perfusion.

9. The method of claim 8 wherein the uppermost location of said medium is at least 2.5 cm above the lowermost location of said medium.

10. The method of claim 8 wherein the uppermost location of said medium is at least 3.2 cm above the lowermost location of said medium.

11. The method of claim 8 wherein the uppermost location of said medium is at least 4.0 cm above the lowermost location of said medium.

12. The method of claim 8 wherein the uppermost location of said medium is at least 5.09 cm above the lowermost location of said medium.

13. The method of claim 8 wherein the uppermost location of said medium is at least 6.0 cm above the lowermost location of said medium.

14. The method of claim 8 wherein the uppermost location of said medium is at least 7.0 cm above the lowermost location of said medium.

15. The method of claim 8 wherein the uppermost location of said medium is at least 8.0 cm above the lowermost location of said medium.

16. The method of claim 8 wherein the uppermost location of said medium is at least 9.0 cm above the lowermost location of said medium.

17. The method of claim 8 wherein the uppermost location of said medium is at least 10.2 cm above the lowermost location of said medium.

18. A method of culturing cells comprising:

adding medium and animal cells into a static cell culture device that is not compartmentalized by a semi-permeable membrane, said device including one or more cell growth surfaces for the culture of animal cells comprised at least in part of a non porous gas permeable material, at least a portion of said non porous gas permeable material is in contact with ambient gas, and

placing said device in a location that includes ambient gas at a composition suitable for animal cell culture, wherein said medium is in contact with said non porous gas permeable material, and the uppermost location of said medium is more than 2.0 cm above the lowermost location of said medium, and said device is not subjected to mixing or perfusion.

19. The method of claim 18 wherein the uppermost location of said medium is at least 2.5 cm above the lowermost location of said medium.

20. The method of claim 18 wherein the uppermost location of said medium is at least 3.2 cm above the lowermost location of said medium.

21. The method of claim 18 wherein the uppermost location of said medium is at least 4.0 cm above the lowermost location of said medium.

22. The method of claim 18 wherein the uppermost location of said medium is at least 5.09 cm above the lowermost location of said medium.

23. The method of claim 18 wherein the uppermost location of said medium is at least 6.0 cm above the lowermost location of said medium.

24. The method of claim 18 wherein said device includes a gas permeable material support in contact with said gas permeable, liquid impermeable material.

25. A method of culturing cells comprising:

adding medium and animal cells into a static cell culture device that is not compartmentalized by a semi-permeable membrane, said device is comprised at least in part of a non porous gas permeable material, at least a portion of said non porous gas permeable material is in contact with ambient gas and is in contact with a gas permeable material support, and

placing said device in a location that includes ambient gas at a composition suitable for animal cell culture, wherein said non porous gas permeable material is in a horizontal plane, and the uppermost location of said medium is elevated more than 2.0 cm beyond the lowermost location of said medium, and said device is not subjected to mixing or perfusion.

26. The method of claim 25 wherein the uppermost location of said medium is elevated at least 2.5 cm beyond the lowermost location of said medium.

27. The method of claim 25 wherein the uppermost location of said medium is elevated at least 3.2 cm beyond the lowermost location of said medium.

28. The method of claim 25 wherein the uppermost location of said medium is elevated at least 4.0 cm beyond the lowermost location of said medium.

29. The method of claim 25 wherein the uppermost location of said medium is elevated at least 5.09 cm beyond the lowermost location of said medium.

30. The method of claim 25 wherein the uppermost location of said medium is elevated at least 6.0 cm beyond the lowermost location of said medium.