Cell carrier, methods of making and use

Patent metadata

Field	Value
Publication number	US-9453196-B2
Application number	US-201113287611-A
Country	US
Kind code	B2
Filing date	Nov 2, 2011
Priority date	Dec 16, 2010
Publication date	Sep 27, 2016
Grant date	Sep 27, 2016

Abstract

A method of culturing adherent cells is provided, wherein the method comprises providing a carrier for growing the cells, comprising one or more surfaces; and one or more relief features/indentations are present on one or more of the surfaces, wherein the carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.012 mm to 0.5 mm; and wherein each of the relief features/indentations has a height above the surfaces in a range from about 2 to 200 μm, and width in a range from about 20 to 200 μm; seeding the cells on the carriers; and growing the cells on the carrier.

First claim

The invention claimed is:

1. A method of culturing adherent cells, comprising:
providing a carrier for growing the cells, comprising a first surface and a second surface opposing the first surface; and one or more relief features on the first surface and the second surface, the relief features on the first surface and the second surface comprising discontinuous protrusions that are positioned away from an edge of the carrier, wherein the carrier has a length in a range from about 0.2 mm to 25 mm, a width in a range from about 0.2 mm to 25 mm, and a height in a range from about 0.012 mm to 0.5 mm; and wherein each of the relief features has a height above the surfaces in a range from about 2 to 200 μm, and width in a range from about 20 to 200 μm and wherein the relief features on the first surface and the second surface comprise discontinuous protrusions that are positioned away from an edge of the carrier.

2. The method of claim 1 , wherein the relief features prevent clumping of two or more carriers while growing the cells.

3. The method of claim 1 , further comprising growing the cells on the carrier in a bioreactor.

4. The method of claim 3 , wherein the bioreactor comprises fluid having a forced convective fluid motion.

5. The method of claim 3 , wherein the bioreactor is a stirred tank bioreactor, a reactor with rocking or rolling motion, a reactor with packed bed or a reactor with fluidized bed.

6. The method of claim 3 , wherein the bioreactor is a stirred tank bioreactor.

7. The method of claim 3 , wherein the bioreactor comprises a disposable housing comprising fluid.

8. The method of claim 7 , further comprising loading the carriers to the disposable housing for growing cells.

9. The method of claim 7 , wherein the disposable housing is a bag, a flask, a tube, a petri dish, or a bottle.

10. The method of claim 7 , wherein the relief features prevent sticking of one or more carriers to one or more walls of the disposable housing.

11. The method of claim 1 , wherein the relief features decrease stacking of two or more carriers.

12. The method of claim 1 , wherein the cells are growing at a temperature in a range of 30° C. to 37° C.

13. The method of claim 1 , wherein the cells are released after growing on the carrier using a mechanical method, a chelant, an enzyme, a thermoresponsive coating, a pH responsive coating, or a combination thereof.

14. The method of claim 13 , wherein the cells are released after growing on the carrier using a thermoresponsive polymer coating.

15. The method of claim 14 , wherein the cells are released at a temperature below lower critical solution temperature (LCST) of the thermoresponsive polymer.

16. The method of claim 1 , wherein the cells are human mesenchymal stromal cells (hMSC), Chinese hamster ovary (CHO) cells, Madin-Darby canine kidney (MDCK) cells, or Vero cells.

17. The method of claim 1 , wherein the relief features disposed on the first surface comprise protrusions having a first shape, and wherein the relief features disposed on the second surface comprise protrusions having a second shape.

18. The method of claim 1 , wherein the relief features disposed on the first surface and the second surface comprise protrusions having rotational symmetry about an axis orthogonal to a plane of the carrier.

19. The method of claim 1 , wherein the relief features comprise profiled post protrusions that narrow as they extend away from the surface of the carrier.

20. The method of claim 1 , wherein the relief features comprise domed protrusions.

21. The method of claim 1 , wherein the relief features comprise cylindrical post protrusions.

22. The method of claim 1 , wherein the relief features disposed on the first surface comprise protrusions oriented along a first axis, and wherein the relief features disposed on the second surface comprise protrusions oriented along a second axis.

23. A method of culturing adherent cells, comprising:
providing a carrier for growing the cells, comprising a first surface and a second surface opposing the first surface; and one or more relief features are present on the first surface and the second surface, wherein the relief features disposed on the first surface comprise protrusions having a first shape, and wherein the relief features disposed on the second surface comprise protrusions having a second shape, wherein the carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.012 mm to 0.5 mm; and wherein each of the relief features has a height above the surfaces in a range from about 2 to 200 μm, and width in a range from about 20 to 200 μm;
seeding the cells on the first surface and the second surface of the carrier;
loading the carrier to a disposable housing comprising fluid;
loading the disposable housing to a bioreactor; and
growing the cells on the carrier in the bioreactor.

24. The method of claim 23 , wherein the disposable housing is a bag, a flask, a tube, a petri dish, or a bottle.

25. The method of claim 23 , wherein the relief features reduce clumping of two or more of the carrier or sticking of one or more carrier to one or more walls of the disposable housing.

26. The method of claim 23 , wherein the fluid have a forced convective fluid motion.

27. The method of claim 23 , wherein the bioreactor is a stirred tank bioreactor, a reactor with rocking or rolling motion, a reactor with packed bed or a reactor with fluidized bed.

28. The method of claim 23 , wherein the bioreactor is a stirred tank bioreactor.

Assignees

• Smith Reginald Donovan
• Miller Scott Michael
• Yoon Yosang
• Rubinsztajn Slawomir
• Susarla Prameela
• Gen Electric

Inventors

• Smith Reginald Donovan
• Miller Scott Michael
• Yoon Yosang
• Rubinsztajn Slawomir
• Susarla Prameela

Classifications

• C12N5/0068 Primary

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12N2535/10

  Patterned coating · CPC title

• C12N2533/30

  Synthetic polymers · CPC title

• C12N2533/12

  Glass · CPC title