IP Library › Granted Patent US 8,748,120
Granted Patent B2
US 8,748,120 · App. 13/953,225 · Granted Jun 10, 2014

Method of selecting a surface chemistry for culturing a given cell line

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Quick Facts
Patent No.
US 8,748,120
App. No.
13/953,225
Granted
Jun 10, 2014
Kind
B2
Abstract

In one embodiment of the invention, there is provided a method of selecting a surface chemistry for culturing a given cell line, including the steps of: (i) providing a first substrate having a plasma polymerized surface with first and second regions, said first region having a first concentration of carboxylic acid groups on said plasma polymerized surface and said second region having a second concentration of carboxylic acid groups on said plasma polymerized surface, wherein said first and second concentrations are different; (ii) culturing cells from said cell line on said plasma polymerized surface in each said region; (iii) observing activity of said cultured cells in each said region, said activity selected from the group consisting of alkaline phosphatase activity of the cultured cells, cell growth of the cultured cells, attachment of the cultured cells, and cell spreading of the cultured cells; and iv) selecting, based on a particular observed activity, a secondary substrate for culturing said cells wherein the secondary substrate has a plasma polymerized surface having a constant concentration of carboxylic acid groups thereon which is equal to the concentration of carboxylic acid groups in the region on said first substrate wherein the cells exhibit said particular observed activity.

Claims (24)

1. A method of selecting a surface chemistry for culturing a given cell line, comprising the steps of:

i) providing a first substrate comprising a plasma polymerized surface having first and second regions, said first region comprising a first concentration of carboxylic acid groups on said plasma polymerized surface and said second region comprising a second concentration of carboxylic acid groups on said plasma polymerized surface, wherein said first and second concentrations are different;

ii) culturing cells from said cell line on said plasma polymerized surface in each said region;

iii) observing activity of said cultured cells in each said region, said activity selected from the group consisting of alkaline phosphatase activity of the cultured cells, cell growth of the cultured cells, attachment of the cultured cells, and cell spreading of the cultured cells; and

iv) selecting, based on a particular observed activity, a secondary substrate for culturing said cells wherein the secondary substrate comprises a plasma polymerized surface having a constant concentration of carboxylic acid groups thereon which is equal to the concentration of carboxylic acid groups in the region on said first substrate wherein the cells exhibit said particular observed activity.

2. The method of claim 1 , wherein said particular observed activity is the level of alkaline phosphatase activity of the cultured cells.

3. The method of claim 2 , wherein said level of alkaline phosphatase activity is observed through alkaline phosphatase staining of said cultured cells.

4. The method of claim 1 , wherein said particular observed activity is the degree of cell growth of said cultured cells.

5. The method of claim 1 , wherein said particular observed activity is the degree of attachment of said cultured cells.

6. The method of claim 1 , wherein said particular observed activity is the level of cell spreading of said cultured cells.

7. The method of claim 1 , wherein said first region comprises a concentration of carboxylic acid groups from 3% to 33%.

8. The method of claim 1 , wherein said second region comprises a concentration of carboxylic acid groups from 3% to 33%.

9. The method of claim 1 , wherein said first region comprises a concentration of carboxylic acid groups from 14% to 20%.

10. The method of claim 1 , wherein said second region comprises a concentration of carboxylic acid groups from 14% to 20%.

11. The method of claim 1 , wherein said first region comprises a concentration of carboxylic acid groups from 2% to 11%.

12. The method of claim 1 , wherein said second region comprises a concentration of carboxylic acid groups from 2% to 11%.

13. The method of claim 1 , wherein said cell line comprises pluripotent cells.

14. The method of claim 13 , wherein said pluripotent cells comprise embryonic stem cells.

15. The method of claim 14 , wherein said embryonic stem cells comprise human embryonic stem cells.

16. The method of claim 1 , wherein said step of culturing cells comprises culturing cells in an animal component free media.

17. The method of claim 1 , wherein said step of providing said plasma polymerized surface having first and second regions comprises depositing a known amount of polymer on said surface.

18. The method of claim 1 , wherein said plasma polymerized surface comprises plasma deposited gradients of octadiene to acrylic acid.

19. The method of claim 18 , wherein said step of providing said plasma polymerized surface comprises the step of depositing a known amount of octadiene and acrylic acid in each of said first and second regions.

20. The method of claim 19 , wherein the rate at which the amount of octadiene and acrylic acid is deposited each of said first and second regions is variable.