Cell carrier, associated methods for making cell carrier and culturing cells using the same

Patent metadata

Field	Value
Publication number	US-9518249-B2
Application number	US-201313839049-A
Country	US
Kind code	B2
Filing date	Mar 15, 2013
Priority date	Dec 16, 2010
Publication date	Dec 13, 2016
Grant date	Dec 13, 2016

Abstract

A carrier for expansion of pluripotent stem cells is provided, wherein the carrier comprises a substrate comprising one or more outer surfaces, wherein the one or more outer surfaces are modified with gas plasma treatment, and one or more structured indentations on one or more of the outer surfaces. 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 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to about 0.5 mm. A method of making the carrier, and culturing stromal cells using the same carrier are also provided.

First claim

The invention claimed is:

1. A carrier for expansion of pluripotent stem cells, comprising:
a planar substrate disc comprising a first surface and a second surface opposing the first surface and a plurality of structured indentations formed in the first surface and the second surface;
wherein the first surface and the second surface are modified with gas plasma treatment, and
wherein the planar substrate disc 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 0.05 mm to 1.2 mm, wherein the height extends from a highest point of the first surface through the planar substrate disc and to a highest point of the second surface and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to 0.5 mm and wherein a first structured indentation of the plurality of structured indentations is aligned with a second structured indentation of the plurality of structured indentations in the planar substrate disc such that respective bottoms of the first structured indentation and the second structured indentation are separated by a portion of the planar substrate disc.

2. The carrier of claim 1 , wherein the first surface and the second surface are configured for expansion of pluripotent stem cells.

3. The carrier of claim 1 , wherein the gas plasma treatment comprises plasma treatment with one or more gases comprising oxygen, nitrogen, amonia, carbon dioxide or combinations thereof.

4. The carrier of claim 1 , wherein the gas plasma treatment comprises oxygen plasma treatment.

5. The carrier of claim 1 , further comprising a biomolecular coating disposed on one or more of the first surface or the second surface.

6. The carrier of claim 5 , wherein the biomolecular coating comprises proteins or peptides.

7. The carrier of claim 5 , wherein the biomolecular coating comprises proteins.

8. The carrier of claim 7 , wherein the proteins comprise collagen, vitronectin, fibronectin, laminin, recombinant collagen, recombinant vitronectin, recombinant e-cadherin or combinations thereof.

9. The carrier of claim 5 , wherein the biomolecular coating comprises synthetic peptides.

10. The carrier of claim 5 , wherein the biomolecular coating comprises extracellular matrix (ECM) proteins, proteoglycans, factors derived from a mouse sarcoma cell line, or combinations thereof.

11. The carrier of claim 5 , wherein the biomolecular coating forms a cytophilic surface.

12. The carrier of claim 1 , wherein each of the structured indentations comprises a cross sectional profile of polygonal, circular, or elliptical shape.

13. The carrier of claim 1 , wherein each of the structured indentations has an aspect ratio in a range from about 0.1 to about 1.5.

14. The carrier of claim 1 , wherein the carrier is made of glass, a polymer, a ceramic, a metal, or combinations thereof.

15. The carrier of claim 1 , wherein the carrier is made of dextran, silicone, polyester, polycarbonate, polyamide, polyurethane, olefin polymer, or polyacrylate polymer.

16. The carrier of claim 1 , wherein the carrier is made of polystyrene.

17. The carrier of claim 1 , having a perimeter that is triangular, rectangular, square, pentagonal, hexagonal, circular, or elliptical.

18. A cell culture system comprising at least one of the carrier of claim 1 .

19. The cell culture system of claim 18 , wherein the cell culture system is a bioreactor.

20. The cell culture system of claim 19 , wherein the bioreactor comprises a fluid having a forced convective fluid motion.

21. The cell culture system of claim 20 , wherein the bioreactor is a stirred tank bioreactor, or a reactor with rocking or rolling motion.

22. A kit for culturing cells, comprising a disposable housing pre-loaded with the carrier of claim 1 .

23. The kit of claim 22 , wherein the disposable housing comprises a bag, a flask, a tube, a petridish, or a bottle.

24. A carrier for expansion of pluripotent stem cells, comprising:
a planar substrate comprising a first surface and a second surface opposing the first surface and a plurality of structured indentations on the first surface and the second surface;
wherein the first surface and the second surface comprise a biomolecular coating,
wherein the planar substrate 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 0.05 mm to 1.2 mm, wherein the height extends from a highest point of the first surface through the planar substrate and to a highest point of the second surface, and wherein each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to 0.5 mm and wherein a first structured indentation of the plurality of structured indentations is aligned with a second structured indentation of the plurality of structured indentations in the planar substrate such that respective bottoms of the first structured indentation and the second structured indentation are separated by a portion of the planar substrate; and
a plurality of pluripotent stem cells disposed in a least one of the plurality of structured indentations.

25. The carrier of claim 24 , wherein the biomolecular coating comprises proteins or peptides.

26. A carrier for expansion of pluripotent stem cells, comprising:
a planar substrate comprising a first surface and a second surface opposing the first surface modified with one or more of corona discharge treatment, gas plasma treatment, or chemical functionalization to form modified surfaces and a plurality of structured indentations on the first surface and the second surface;
a biomolecular coating disposed on the first or second surface,
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 0.05 mm to 1.2 mm wherein the height extends from a highest point of the first surface through the planar substrate and to a highest point of the second surface, and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm, and a depth in a range from about 0.025 mm to about 0.5 mm and wherein a first structured indentation of the plurality of structured indentations is aligned with a second structured indentation of the plurality of structured indentations in the planar substrate such that respective bottoms of the first structured indentation and the second structured indentation are separated by a portion of the planar substrate; and
a plurality of pluripotent stem cells disposed in at least one of the plurality of indentations.

27. The carrier of claim 26 , wherein the gas plasma treatment comprises plasma treatment with one or more of the gases comprising oxygen, nitrogen, amonia, carbon dioxide, or combinations thereof.

Assignees

• Gen Electric

Inventors

• Davis Brian Michael
• Loghin Evelina Roxana
• Conway Kenneth Roger
• Burns Andrew Arthur Paul
• Gascoyne David Gilles

Classifications

• C12N2501/999

  Small molecules not provided for elsewhere · CPC title

• C12N5/0068 Primary

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

• Y10T156/1039

  Surface deformation only of sandwich or lamina [e.g., embossed panels] · CPC title

• C12N2533/90

  Substrates of biological origin, e.g. extracellular matrix, decellularised tissue · CPC title

• C12N2537/00

  Supports and/or coatings for cell culture characterised by physical or chemical treatment · CPC title