Cell culturing method, cell culturing apparatus and kit comprising a porous polyimide film

• US 9,868,933 B2
• Filed: 07/25/2014
• Issued: 01/16/2018
• Est. Priority Date: 07/26/2013
• Status: Active Grant

• Current Assignee: UBE Corporation
• Original Assignee: UBE Industries Limited (UBE Corporation)
• Inventors: Hagihara, Masahiko, Kawaguchi, Tetsuo, Baba, Kousuke, Shimizu, Motohisa
• Primary Examiner(s): Barron, Sean C
• Application Number:
  US14/907,432
• Publication Number:
  US 20160168560A1
• Time in Patent Office: 1,271 Days
• Field of Search: 435180, 4352521, 4352525, 4352528, 4352551, 435348, 435395, 435420
• CPC Class Codes: C08L 79/08 Polyimides; Polyester-imides; Polyamide-imides; Polyamide acids or similar polyimide precursors
• CPC Class Codes: C12M 25/02 Membranes; Filters filters or filtration in general B01D24/00-B01D41/00
• CPC Class Codes: C12N 1/16 Yeasts; Culture media therefor
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12N 11/089 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

The present invention relates to a cell culturing method and cell culturing apparatus and kit for use therewith. The method includes applying cells on a porous polyimide film and culturing. One embodiment includes a process for sowing cells on the surface of the film. Another embodiment includes a process for placing a cell suspension on the dried surface of the film and leaving the film undisturbed, moving the film to promote liquid efflux, or stimulating a portion of the surface to entangle the cell suspension into the film, and then retaining the cells in suspension inside the film while allowing moisture to flow out. Another embodiment includes a process for moistening one or both surfaces of the film with a cell culture solution or sterilized liquid, loading a cell suspension on the moistened film, retaining the cells in suspension inside the film, and allowing moisture to flow out.

Claims

1. A cell culturing method that includes applying cells to a porous polyimide film and culturing them, wherein

the porous polyimide film has a three-layer structure consisting of an A-surface layer having a plurality of pores, a B-surface layer having a plurality of pores, and a macro-void layer sandwiched between the two surface layers,

a mean pore size in the A-surface layer is smaller than a mean pore size in the B-surface layer, and

the macro-void layer has a partition bonded to the A-surface layer and the B-surface layer, and a plurality of macro-voids surrounded by the partition, the A-surface layer, and the B-surface layer.

2. The method according to claim 1, including a step of seeding cells on either the A-surface or the B-surface of the porous polyimide film.

3. The method according to claim 1, including a step of:

placing a cell suspension on the dried surface of the porous polyimide film,

allowing the porous polyimide film to stand, or moving the porous polyimide film to promote efflux of the liquid, or stimulating part of the surface to cause absorption of the cell suspension into the film, and

retaining the cells in the cell suspension inside the film and allowing the water to flow out.

4. The method according to claim 1, including a step of:

wetting one or both sides of the porous polyimide film with a cell culture medium or a sterilized liquid,

loading a cell suspension into the wetted porous polyimide film, and

retaining the cells in the cell suspension inside the film and allowing the water to flow out.

5. The method according to claim 4, wherein the viable cells are retained in the porous polyimide film, and the dead cells are allowed to flow out together with the water.

6. The method according to claim 4, wherein the sterilized liquid is sterilized water or a sterilized buffering solution.

7. The method according to claim 1, including the aspect that the cell culture medium, cells and one or more porous polyimide films are placed in a cell culturing vessel, wherein the porous polyimide film is in a suspended state in the cell culture medium.

8. The method according to claim 7, wherein two or more fragments of the porous polyimide film are used.

9. The method according to claim 7, wherein the cells spontaneously adhere to the porous polyimide film.

10. The method according to claim 1, wherein the porous polyimide film is:

i) folded,

ii) wound into a roll,

iii) connected as sheets or fragments by a filamentous structure, or

iv) bound into a rope,

for suspension or fixing in the cell culture medium in the cell culturing vessel.

11. The method according to claim 10, wherein the cells spontaneously adhere to the porous polyimide film.

12. The method according to claim 1, including using two or more porous polyimide films layered either above and below or left and right in the cell culture medium.

13. The method according to claim 1, wherein the cells grow and proliferate on the surface of and inside the porous polyimide film.

14. The method according to claim 1, wherein the cells are selected from the group consisting of animal cells, insect cells, plant cells, yeast cells and bacteria.

15. The method according to claim 14, wherein the animal cells are cells derived from an animal belonging to the subphylum Vertebrata.

16. The method according to claim 14, wherein the bacteria are selected from the group consisting of lactic acid bacteria, E. coli, Bacillus subtilis and cyanobacteria.

17. The method according to claim 1,

wherein the cells are selected from the group consisting of pluripotent stem cells, tissue stem cells, somatic cells and germ cells.

18. The method according to claim 1, wherein the cells are selected from the group consisting of sarcoma cells, established cell lines and transformants.

19. The method according to claim 1, wherein the porous polyimide film is a porous polyimide film including a polyimide obtained from a tetracarboxylic dianhydride and a diamine.