Cell carrier, associated methods for making cell carrier and culturing cells using the same
View Patent ↗A carrier for expansion of induced 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 induced pluripotent stem cells using the same carrier are also provided.
1. A carrier for expansion of induced pluripotent stem cells, wherein the carrier is configured for suspension in a bioreactor, and for protecting the cells from exposure to hydrodynamic shear generated by fluid motion in the bioreactor, comprising:
a substrate comprising two outer surfaces, wherein one or more structured indentations are present on the two outer surfaces;
wherein the two outer surfaces are modified with gas plasma treatment, the carrier has a length of about 6.5 mm, a width of about 6.5 mm, and a height of about 0.5 mm and each of the structured indentations has a major axis of about 0.45 mm, a minor axis of about 0.45 mm and a depth of about 0.2 mm.
2. The carrier of claim 1 , wherein the gas plasma treatment comprises plasma treatment with one or more of the gases comprising oxygen, nitrogen, ammonia, carbon dioxide, nitrous oxide or combinations thereof.
3. The carrier of claim 1 , wherein the gas plasma treatment comprises oxygen plasma treatment.
4. The carrier of claim 1 , wherein the gas plasma treated carrier comprises a water contact angle between 10 and 70 degrees.
5. The carrier of claim 1 , further comprising a biomolecular coating disposed on the two outer surfaces.
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, e-cadherin, recombinant 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 has a cross sectional profile of polygonal, circular, or elliptical shape.
13. The carrier of claim 1 , wherein the carrier is made of a glass, polymer, ceramic, metal or combination thereof.
14. The carrier of claim 1 , wherein the carrier is made of dextran, silicone, polyester, polycarbonate, polyamide, polyurethane, olefin polymer, or polyacrylate polymer.
15. The carrier of claim 1 , wherein the carrier is made of polystyrene.
16. The carrier of claim 1 , wherein the carrier is made of a material having a density between 1.0 and 1.4.
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 one or more of the carriers 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 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 petri dish, or a bottle.
24. A carrier for expansion of induced pluripotent stem cells, wherein the carrier is configured for suspension in a bioreactor, and for protecting the cells from exposure to hydrodynamic shear generated by fluid motion in a bioreactor, comprising:
a substrate comprising two outer surfaces, wherein one or more structured indentations are present on the two outer surfaces;
wherein the two outer surfaces comprise a biomolecular coating, the carrier has a length of about 6.5 mm, a width of about 6.5 mm, and a height of about 0.5 mm and each of the structured indentations has a major axis of about 0.45 mm, a minor axis of about 0.45 mm and a depth of about 0.2 mm.
25. The carrier of claim 24 , wherein the biomolecular coating comprises proteins or peptides.
26. A carrier for expansion of induced pluripotent stem cells, wherein the carrier is configured for suspension in a bioreactor, and for protecting the cells from exposure to hydrodynamic shear generated by fluid motion in a bioreactor, comprising:
a substrate comprising two outer surfaces, wherein one or more structured indentations are present on the two outer surfaces; wherein the two outer surfaces are modified with one or more of corona discharge treatment, gas plasma treatment, or chemical functionalization to form modified surfaces, and
a biomolecular coating disposed on the two modified surfaces, the carrier has a length of about 6.5 mm, a width of about 6.5 mm, and a height of about 0.5 mm and each of the structured indentations has a major axis of about 0.45 mm, a minor axis of about 0.45 mm and a depth of about 0.2 mm.
27. The carrier of claim 26 , wherein the gas plasma treatment comprises plasma treatment with one or more of the gases comprising oxygen, nitrogen, ammonia, carbon dioxide or combinations thereof.