Synthetic Surfaces for Culturing Stem Cell Derived Oligodendrocyte Progenitor Cells

• US 20090191626A1
• Filed: 01/29/2009
• Published: 07/30/2009
• Est. Priority Date: 01/30/2008
• Status: Active Grant

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Inventors: Zhou, Yue, Shogbon, Christopher Bankole, Brandenberger, Ralph
• Granted Patent: US 8,513,009 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0622 Glial cells, e.g. astrocytes, oligodendrocytes; Schwann cells
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

Synthetic surfaces suitable for culturing stem cell derived oligodendrocyte progenitor cells contain acrylate polymers formed from one or more acrylate monomers. The acrylate surfaces, in many cases, are suitable for culturing stem cell derived oligodendrocyte progenitor cells in chemically defined media.

Claims

1. A cell culture article for culturing oligodendrocyte progenitor cells in a chemically defined medium, comprising:

a substrate having a surface;

a polymer material disposed on the surface, wherein the polymer material comprises

(i) a homopolymer formed from tetra(ethylene glycol) diacrylate or

(ii) a copolymer formed from

tetra(ethylene glycol) diacrylate and neopentyl glycol ethoxylate diacrylate,

glycerol dimethacrylate and tetra(ethylene glycol) diacrylate,

glycerol dimethacrylate and tri(ethylene glycol) dimethacrylate,

glycerol dimethacrylate and di(ethylene glycol) dimethacrylate,

glycerol dimethacrylate and tetraethylene glycol dimethacrylate,

glycerol dimethacrylate and 1,6-hexanediol propoxylate diacrylate,

glycerol dimethacrylate and 1,6-hexanediol ethoxylate diacrylate,

tri(ethylene glycol) dimethacrylate and trimethylolpropane triacrylate,

1,4-butanediol dimethacrylate and 1,9 nonanediol diacrylate,

1,6-hexanediol diacrylate and tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate,

1,6-hexanediol diacrylate and 1,6-hexanediol ethoxylate diacrylate,

1,6-hexanediol diacrylate and neopentyl glycol ethoxylate diacrylate,

neopentyl glycol propoxylate (1 PO/OH) diacrylate and neopentyl glycol ethoxylate diacrylate,

di(ethylene glycol) dimethacrylate and neopentyl glycol diacrylate,

di(ethylene glycol) dimethacrylate and tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate,

tetra(ethylene glycol) dimethacrylate and 1,4-butanediol dimethacrylate,

tetra(ethylene glycol) dimethacrylate and neopentyl glycol ethoxylate diacrylate,

1,6-hexanediol propoxylate diacrylate and neopentyl glycol ethoxylate diacrylate,

neopentyl glycol diacrylate and tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate,

neopentyl glycol diacrylate and 1,6-hexanediol ethoxylate diacrylate,

neopentyl glycol diacrylate and poly(propylene glycol) diacrylate,

trimethylolpropane ethoxylate (1 EO/OH) methyl diacrylate and

2,2,3,3,4,4,5,5octafluoro1,6hexanediol diacrylate,

neopentyl glycol ethoxylate diacrylate and di(ethylene glycol) dimethacrylate,

trimethylolpropane triacrylate and 1,4-butanediol dimethacrylate,

trimethylolpropane triacrylate and di(ethylene glycol) dimethacrylate;

trimethylolpropane triacrylate and neopentyl glycol diacrylate,

trimethylolpropane triacrylate and neopentyl glycol dimethacrylate,

2,2,3,3,4,4,5,5octafluoro1,6hexanediol diacrylate and tetra(ethylene glycol) dimethacrylate,

2,2,3,3,4,4,5,5octafluoro1,6hexanediol diacrylate and neopentyl glycol diacrylate, or

poly(propylene glycol) diacrylate and glycerol 1,3-diglycerolate diacrylate.

2. The article of claim 1, wherein the article is produced by a process of:

diluting one or more monomers in a volatile organic solvent;

depositing the diluted monomers on the substrate surface of the article;

evaporating substantially all of the solvent; and

polymerizing the monomers to form the polymer layer attached to the surface.

3. The article of claim 2 wherein the volatile solvent is ethanol.

4. The article of claim 1 wherein the polymer material disposed on the surface of the article has a surface area greater than about 5 mm².

5. The article of claim 1, further comprising a well, wherein the surface of the substrate on which the polymer material is disposed is in the well.

6. The article of claim 5, wherein the article is selected from the group consisting of a multi-well plate, a petri dish, a beaker, a tube, and a flask.

7. A culture of oligodendrocyte progenitor cells, comprising:

an article comprising a polymer material disposed on a surface;

the oligodendrocyte progenitor cells disposed on the polymer material; and

a culture medium in which the oligodendrocyte progenitor cells are cultured,

wherein the polymer material comprises

(i) a homopolymer formed from tetra(ethylene glycol) diacrylate or

(ii) a copolymer formed from

tetra(ethylene glycol) diacrylate and neopentyl glycol ethoxylate diacrylate,

glycerol dimethacrylate and tetra(ethylene glycol) diacrylate,

glycerol dimethacrylate and tri(ethylene glycol) dimethacrylate,

glycerol dimethacrylate and di(ethylene glycol) dimethacrylate,

glycerol dimethacrylate and tetraethylene glycol dimethacrylate,

glycerol dimethacrylate and 1,6-hexanediol propoxylate diacrylate,

glycerol dimethacrylate and 1,6-hexanediol ethoxylate diacrylate,

tri(ethylene glycol) dimethacrylate and trimethylolpropane triacrylate,

1,4-butanediol dimethacrylate and 1,9 nonanediol diacrylate,

1,6-hexanediol diacrylate and tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate,

1,6-hexanediol diacrylate and 1,6-hexanediol ethoxylate diacrylate,

1,6-hexanediol diacrylate and neopentyl glycol ethoxylate diacrylate,

neopentyl glycol propoxylate (lPO/OH) diacrylate and neopentyl glycol ethoxylate diacrylate,

di(ethylene glycol) dimethacrylate and neopentyl glycol diacrylate,

di(ethylene glycol) dimethacrylate and tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate,

tetra(ethylene glycol) dimethacrylate and 1,4-butanediol dimethacrylate,

tetra(ethylene glycol) dimethacrylate and neopentyl glycol ethoxylate diacrylate,

1,6-hexanediol propoxylate diacrylate and neopentyl glycol ethoxylate diacrylate,

neopentyl glycol diacrylate and tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate,

neopentyl glycol diacrylate and 1,6-hexanediol ethoxylate diacrylate,

neopentyl glycol diacrylate and poly(propylene glycol) diacrylate,

trimethylolpropane ethoxylate (1 EO/OH) methyl diacrylate and 2,2,3,3,4,4,5,5octafluoro1,6hexanediol diacrylate,

neopentyl glycol ethoxylate diacrylate and di(ethylene glycol) dimethacrylate,

trimethylolpropane triacrylate and 1,4-butanediol dimethacrylate,

trimethylolpropane triacrylate and di(ethylene glycol) dimethacrylate;

trimethylolpropane triacrylate and neopentyl glycol diacrylate,

trimethylolpropane triacrylate and neopentyl glycol dimethacrylate,

2,2,3,3,4,4,5,5octafluoro1,6hexanediol diacrylate and tetra(ethylene glycol) dimethacrylate,

2,2,3,3,4,4,5,5octafluoro1,6hexanediol diacrylate and neopentyl glycol diacrylate, or

poly(propylene glycol) diacrylate and glycerol 1,3-diglycerolate diacrylate.

8. The culture of claim 7, wherein the oligodendrocyte progenitor cells are stem cell derived oligodendrocyte progenitor cells.

9. The culture of claim 7, wherein the oligodendrocyte progenitor cells are human embryonic stem cell derived oligodendrocyte progenitor cells.

10. A method for culturing oligodendrocyte progenitor cells, comprising:

depositing a suspension comprising the oligodendrocyte progenitor cells on a polymer material; and

culturing the deposited oligodendrocyte progenitor cells in a cell culture medium,

wherein the polymer material comprises

(i) a homopolymer formed from tetra(ethylene glycol) diacrylate or

(ii) a copolymer formed from

tetra(ethylene glycol) diacrylate and neopentyl glycol ethoxylate diacrylate,

glycerol dimethacrylate and tetra(ethylene glycol) diacrylate,

glycerol dimethacrylate and tri(ethylene glycol) dimethacrylate,

glycerol dimethacrylate and di(ethylene glycol) dimethacrylate,

glycerol dimethacrylate and tetraethylene glycol dimethacrylate,

glycerol dimethacrylate and 1,6-hexanediol propoxylate diacrylate,

glycerol dimethacrylate and 1,6-hexanediol ethoxylate diacrylate,

tri(ethylene glycol) dimethacrylate and trimethylolpropane triacrylate,

1,4-butanediol dimethacrylate and 1,9 nonanediol diacrylate,

1,6-hexanediol diacrylate and tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate,

1,6-hexanediol diacrylate and 1,6-hexanediol ethoxylate diacrylate,

1,6-hexanediol diacrylate and neopentyl glycol ethoxylate diacrylate,

neopentyl glycol propoxylate (1 PO/OH) diacrylate and neopentyl glycol ethoxylate diacrylate,

di(ethylene glycol) dimethacrylate and neopentyl glycol diacrylate,

di(ethylene glycol) dimethacrylate and tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate,

tetra(ethylene glycol) dimethacrylate and 1,4-butanediol dimethacrylate,

tetra(ethylene glycol) dimethacrylate and neopentyl glycol ethoxylate diacrylate,

1,6-hexanediol propoxylate diacrylate and neopentyl glycol ethoxylate diacrylate,

neopentyl glycol diacrylate and tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate,

neopentyl glycol diacrylate and 1,6-hexanediol ethoxylate diacrylate,

neopentyl glycol diacrylate and poly(propylene glycol) diacrylate,

trimethylolpropane ethoxylate (1 EO/OH) methyl diacrylate and 2,2,3,3,4,4,5,5octafluoro1,6hexanediol diacrylate,

neopentyl glycol ethoxylate diacrylate and di(ethylene glycol) dimethacrylate,

trimethylolpropane triacrylate and 1,4-butanediol dimethacrylate,

trimethylolpropane triacrylate and di(ethylene glycol) dimethacrylate;

trimethylolpropane triacrylate and neopentyl glycol diacrylate,

trimethylolpropane triacrylate and neopentyl glycol dimethacrylate,

2,2,3,3,4,4,5,5octafluoro1,6hexanediol diacrylate and tetra(ethylene glycol) dimethacrylate,

2,2,3,3,4,4,5,5octafluoro1,6hexanediol diacrylate and neopentyl glycol diacrylate, or

poly(propylene glycol) diacrylate and glycerol 1,3-diglycerolate diacrylate.

11. The method of claim 10, wherein the cell culture medium is a chemically defined medium.

12. The method of claim 10, wherein the oligodendrocyte progenitor cells are stem cell derived oligodendrocyte progenitor cells.

13. The method of claim 10, wherein the oligodendrocyte progenitor cells are human embryonic stem cell derived oligodendrocyte progenitor cells.