Cell culture apparatus and methods

• US 20050221486A1
• Filed: 03/24/2005
• Published: 10/06/2005
• Est. Priority Date: 03/25/2004
• Status: Abandoned Application

• Current Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Original Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Inventors: Kadarusman, Judith, Lin, WengLong Roy, Bhatia, Ravinder, McLaughlin, John
• Application Number:
  US11/087,801
• Publication Number:
  US 20050221486A1
• Time in Patent Office: Days
• US Class Current: 435/404
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 5/0018 Culture media for cell or tissue culture media for specific animal cell type C12N5/06

Abstract

A method for culturing cells in a plastic culture vessel is provided. The method is suitable for use when it is desirable to have a small hydrophobic molecule present in the cell culture media at some point during incubation.

Claims

1. A method for culturing cells in a plastic culture vessel comprising the steps of:

(a) providing a plastic culture vessel constructed from a fluoropolymer;

(b) introducing cell culture media into the cell culturing vessel;

(c) introducing a small hydrophobic molecule into the cell culture media;

(d) inoculating the cell culture media with cells; and

(e) incubating the cell culture under suitable conditions for cell growth.

2. The method of claim 1, wherein the step of introducing a small hydrophobic molecule into the cell culture media comprises exogenous addition of the small hydrophobic molecule to the cell culture media.

3. The method of claim 2, wherein the small hydrophobic molecule is selected from the group consisting of:

(a) a small hydrophobic molecule which is important for cell function; and

(b) a small hydrophobic molecule added to the cell culture media to evaluate an effect of the molecule on cell function.

4. The method of claim 1, wherein the step of introducing the small hydrophobic molecule into the cell culture comprises endogenous production of the small hydrophobic molecule by the cells in the culture.

5. The method of claim 1, wherein the small hydrophobic molecule has a molecular weight of less than about 1000 g/mol.

6. The method of claim 1, wherein the small hydrophobic molecule has a molecular weight of less than about 500 g/mol.

7. The method of claim 1, wherein the small hydrophobic molecule comprises a terpene.

8. The method of claim 1, wherein the small molecule comprises cholesterol.

9. The method of claim 7, wherein the terpene is a steroid hormone.

10. The method of claim 9, wherein the steroid hormone is selected from the group consisting of cholesterol, squalene, cortisol, estradiol, progesterone and testosterone.

11. The method of claim 7, wherein the terpene comprises paclitaxel.

12. The method of claim 1, wherein the fluoropolymer is a homopolymer.

13. The method of claim 1, wherein the fluoropolymer is a copolymer.

14. The method of claim 1, wherein the fluoropolymer is selected from the group consisting of:

polychlorotrifluoroethylene (PCTFE), ethylene-tetrafluoroethylene copolymer (ETFE), fluorinated ethylene-propylene copolymer (FEP), Polytetrafluoroethylene (PTFE), polyvinylidene fluoride (PVDF), perfluoroalkyltetrafluoroethylene copolymer (PFA), tetrafluoroethylene and perfluoromethyl vinyl-ether copolymer (MFA), chlorotrifluoroethylene-vinylidene fluoride copolymer (CTFE/VDF), ethylene-chlorotrifluoroethylene copolymer (ECTFE), polyvinyl fluoride (PVF), and tetrafluoroethylene-hexafluoropropylene copolymer (TFE/HFP).

15. The method of claim 1, wherein the fluoropolymer comprises fluorinated ethylene-propylene copolymer (FEP).

16. The method of claim 1, wherein the cell culture vessel is a cell culture bag.

17. The method of claim 1, wherein the cell culture is selected from the group consisting of:

(a) a small-scale cell culture having a volume of less than about 20 L;

(b) a small-scale cell culture having a volume of less than about 1 L;

(c) a medium-scale cell culture having a volume of less than about 1 L and 500L; and

(d) a large-scale cell culture having a volume up to about 20,000 L.

18. The method of claim 1, wherein the cells are selected from the group consisting of:

animal, insect, bacterial, fungal and plant cells.

19. The method of claim 18, wherein the animal cells comprise mammalian cells.

20. The method of claim 19, wherein the mammalian cell is selected from the group consisting of human, mouse, hamster, monkey, and hybridoma cells.

21. The method of claim 20, wherein human cells are selected from the group consisting of 293, WI38, PER.C6 and Bowes melanoma cells.

22. The method of claim 20, wherein mouse cells are selected from the group consisting of 3T3, NS0, NS1, and Sp2/0.

23. The method of claim 20, wherein hamster cells are selected from the group consisting of CHO, and BHK.

24. The method of claim 20, wherein monkey cells are selected from the group consisting of COS, FRhL, and Vero.

25. The method of claim 18, wherein bacterial cells are selected from the group consisting of E. coli, Streptomyces and Salmonella typhimurium cells.

26. The method of claim 18, wherein fungal cells comprise yeast cells selected from the group consisting of Saccharomyces cerevisiae and Pichia pastoris.

27. The method of claim 26, wherein fungal cells comprise Aspergillus spp.

28. The method of claim 18, wherein insect cells are selected from the group consisting of Drosophila S2 and Spodoptera Sf9 and Sf21 cells.

29. The method of claim 1, wherein cells comprise cholesterol dependent NS0 cells.

30. The method of claim 29, wherein the small hydrophobic molecule comprises cholesterol.

31. A method for culturing cholesterol dependent NS0 cells in a plastic culture vessel comprising the steps of:

(a) providing a plastic culture vessel constructed from a fluoropolymer;

(b) introducing cell culture media into the cell culturing vessel;

(c) inoculating the cell culture media with NS0 cells;

(d) introducing cholesterol into the cell culture media; and

(e) incubating the cell culture under suitable conditions for cell growth.

32. The method of claim 31, wherein the fluoropolymer is selected from the group consisting of:

polychlorotrifluoroethylene (PCTFE), ethylene-tetrafluoroethylene copolymer (ETFE), fluorinated ethylene-propylene copolymer (FEP), Polytetrafluoroethylene (PTFE), polyvinylidene fluoride (PVDF), perfluoroalkyltetrafluoroethylene copolymer (PFA), tetrafluoroethylene and perfluoromethyl vinyl-ether copolymer (MFA), chlorotrifluoroethylene-vinylidene fluoride copolymer (CTFE/VDF), ethylene-chlorotrifluoroethylene copolymer (ECTFE), polyvinyl fluoride (PVF), and tetrafluoroethylene-hexafluoropropylene copolymer (TFE/HFP).

33. The method of claim 31, wherein the fluoropolymer comprises fluorinated ethylene-propylene copolymer (FEP).

34. The method of claim 31, wherein the plastic culture vessel comprises a cell culture bag.

35. A method for culturing cells in a plastic culture vessel comprising the steps of:

(a) providing a plastic culture vessel constructed from a vinyl acetate copolymer;

(b) introducing cell culture media into the cell culturing vessel;

(c) introducing a small hydrophobic molecule into the cell culture media;

(d) inoculating the cell culture media with cells; and

(e) incubating the cell culture under suitable conditions for cell growth.

36. The method of claim 35, wherein the vinyl acetate copolymer comprises ethylene vinyl acetate.