Synthetic attachment medium for cell culture

Patent metadata

Field	Value
Publication number	US-11046931-B2
Application number	US-201916371252-A
Country	US
Kind code	B2
Filing date	Apr 1, 2019
Priority date	Feb 2, 2012
Publication date	Jun 29, 2021
Grant date	Jun 29, 2021

Abstract

An aqueous cell culture medium composition includes an aqueous cell culture solution configured to support the culture of mammalian cells. The composition further includes a synthetic polymer conjugated to a polypeptide dissolved in the aqueous cell culture solution. The synthetic polymer conjugated to a polypeptide is configured to attach to the surface of a cell culture article under cell culture conditions. Incubation of the aqueous cell culture medium composition on a cell culture surface under cell culture conditions results is attachment to the surface of the synthetic polymer conjugated to the polypeptide.

First claim

What is claimed is:

1. An aqueous cell culture medium composition comprising:
glucose, at least one amino acid and a synthetic copolymer having a covalently attached polypeptide;

wherein the synthetic copolymer is formed from copolymerization of (i) methacrylic acid conjugated to the polypeptide and (ii) hydroxyethylmethacrylate;
wherein when the aqueous cell culture medium containing the synthetic copolymer is introduced into a cell culture article, the synthetic copolymer attaches to the surface of the cell culture article under cell culture conditions.

2. The aqueous cell culture medium composition of claim 1 , wherein the cell culture medium composition is a chemically defined composition.

3. The aqueous cell culture medium composition of claim 1 wherein synthetic copolymer is introduced to cell culture medium in a dry state.

4. The aqueous cell culture medium composition of claim 1 wherein the synthetic copolymer is added to cell culture media dissolved in water.

5. The aqueous cell culture medium composition of claim 1 , wherein the molar ratio of the methacrylic acid conjugated to the polypeptide and the hydroxyethylmethacrylate is between 1 to 50 and 1 to 1.

6. The aqueous cell culture medium composition of claim 1 wherein the synthetic copolymer is formed from copolymerization of the hydroxyethylmethacrylate and MAA-PEO4-polypeptide, wherein MAA is the methacrylic acid, and PEO is polyethylene oxide.

7. The aqueous cell culture medium composition of claim 1 , wherein the weight percentage of the polypeptide relative to the copolymer conjugated to the polypeptide is greater than 10%.

8. The aqueous cell culture medium composition of claim 1 , wherein the polypeptide is a cell adhesive polypeptide.

9. The aqueous cell culture medium composition of claim 1 , wherein the polypeptide comprises an RGD sequence.

10. The aqueous cell culture medium composition of claim 1 , wherein the polypeptide is a vitronectin polypeptide.

11. The aqueous cell culture medium composition of claim 1 , wherein the polypeptide comprises an amino acid sequence of SEQ ID NO:27.

12. The aqueous cell culture medium composition of claim 1 , wherein the copolymer conjugated to the polypeptide has a molecular weight of between 10 kilodaltons and 1000 kilodaltons.

13. The aqueous cell culture medium composition of claim 1 wherein the copolymer has a linear backbone and is crosslink free, wherein the copolymer having the covalently attached polypeptide is soluble in water at 20° C. or less, and wherein the aqueous solution is substantially free of organic solvents.

14. The aqueous cell culture medium composition of claim 1 , wherein the surface of the cell culture article is formed from a ceramic, glass, plastic, polymer, copolymer, or combination thereof.

15. The aqueous cell culture medium composition of claim 14 , wherein the surface of the cell culture article is formed from soda-lime glass, pyrex glass, vycor glass, quartz glass, silicon, poly(vinyl chloride), poly(vinyl alcohol), poly(methyl methacrylate), poly(vinyl acetate-co-maleic anhydride), poly(dimethylsiloxane) monomethacrylate, cyclic olefin polymers, fluorocarbon polymers, polysytrenes, polypropylene, polyethyleneimine, poly(styrene-co-maleic anhydride), or poly(ethylene-co-acrylic acid).

16. The aqueous cell culture medium composition of claim 14 , wherein the cell culture article is chosen from multi-well plates, jars, petri dishes, flasks, beakers, glass bead microcarriers, polymer bead microcarriers, roller bottles, slides, tubes, cover slips, membranes, hollow fibers, cups, spinner bottles, perfusion chambers, bioreactors, and fermenters.

17. The aqueous cell culture medium composition of claim 16 , wherein the cell culture article is chosen from multi-well plates, petri dishes, flasks, and beakers.

Assignees

• Corning Inc
• Coming Incorporated

Inventors

• Caracci Stephen Joseph
• Henry David
• Kelley Jessica Jo
• Lewis Mark Alan
• Zhou Yue

Classifications

• C12N5/0606

  Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696) · CPC title

• C12N5/0663

  Bone marrow mesenchymal stem cells (BM-MSC) · CPC title

• C12N2533/40

  Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers · CPC title

• C12N5/0068 Primary

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

• C12N2533/30

  Synthetic polymers · CPC title