Cell culture substrate

Patent metadata

Field	Value
Publication number	US-2017191026-A1
Application number	US-201515510556-A
Country	US
Kind code	A1
Filing date	Sep 11, 2015
Priority date	Sep 11, 2014
Publication date	Jul 6, 2017
Grant date	—

Abstract

The invention provides a substrate for cell culture comprising a polymer, wherein the polymer comprises —a homopolymer formed from a monomer of formula (Ia) or (Ib); —a copolymer formed from one or more monomers of formula (Ia) and/or (Ib); or —a copolymer formed from one or more monomers of formula (Ia) and/or (Ib), and comprising HEMA; wherein formula (Ia) comprises: (Formula (Ia)) wherein R1 is a C8-C12 straight or branched chain alkyl or alkenyl group, for example a C8-C10 straight or branched chain alkyl group, which may optionally be substituted; and R2 is selected from H and C1-4 alkyl; and wherein formula (Ib) comprises: (Formula (Ib)) wherein R3 is a 6-12 membered ring, for example a 6-8 membered ring, which is a cycloalkyl, cycloheteroalkyl, aryl or heteroaryl group, and which may optionally be substituted; L is a divalent linker group selected from —NH—, —CH2—, and —O—; R4 is a C1-8 organic group, for example a C1-C6 organic group, wherein this group comprises at least one moiety selected from C═O, NH, NH2, COOH, and C═C.

First claim

1 . A substrate for cell culture comprising a polymer, wherein the polymer comprises
a homopolymer formed from a monomer of formula (Ia) or (Ib); a copolymer formed from one or more monomers of formula (Ia) and/or (Ib); or a copolymer formed from one or more monomers of formula (Ia) and/or (Ib), and comprising HEMA; wherein formula (Ia) comprises:

wherein
R1 is a C8-C12 straight or branched chain alkyl or alkenyl group, for example a C8-C10 straight or branched chain alkyl group, which may optionally be substituted; and
R2 is selected from H and C1-4 alkyl; and
wherein formula (Ib) comprises:

wherein
R3 is a 6-12 membered ring, for example a 6-8 membered ring, which is a cycloalkyl, cycloheteroalkyl, aryl or heteroaryl group, and which may optionally be substituted;
L is a divalent linker group selected from —NH—, —CH 2 —, and —O—;
R4 is a C1-8 organic group, for example a C1-C6 organic group, wherein this group comprises at least one moiety selected from C═O, NH, NH 2 , COOH, and C═C.

2 . The Substrate according to claim 1 , wherein R2 is selected from H and C1 or C2 alkyl.

3 . The substrate according to claim 1 or claim 2 , wherein the R1 group and/or the R3 group is substituted.

4 . The substrate according to any preceding claim, wherein R3 is a cycloheteroalkyl, or heteroaryl group, and wherein the heteroatoms in the ring are selected from O, N, S, SO 2 , P, B, Si, and combinations thereof; or R3 is cycloalkyl or aryl with no heteroatoms in the ring.

5 . The substrate according to any preceding claim, wherein R3 is a 6 membered ring.

6 . The substrate according to any preceding claim, wherein the R4 group is straight chain or branched.

7 . The substrate according to any preceding claim, wherein the R4 group is a C1-C5 organic group, such as a C1-C4 organic group.

8 . The substrate according to any preceding claim, wherein the R4 organic group comprises two or more moieties selected from C═O, NH, NH 2 , COOH, and C═C.

9 . The substrate according to any preceding claim, wherein the R4 group is

where R2 is selected from H and C1-4 alkyl; preferably it is selected from H and C1 or C2 alkyl, for example it may be H or C1 alkyl.

10 . The substrate according to any preceding claim, wherein the monomer of formula (Ia) comprises any of the monomers selected from:
2-Ethylhexyl methacrylate:

Octafluoro-2-hydroxy-6-(trifluoromethyl)heptyl methacrylate:

Dodecafluoroheptyl acrylate:

or
Lauryl methacrylate:

11 . The substrate according to any preceding claim, wherein the monomer of formula (Ib) comprises any of the monomers selected from:
N-(4-Hydroxyphenyl)methacrylamide:

2-Norbornyl methacrylate:

N-Phenylmethacrylamide:

Cyclohexyl methacrylate:

Tris[2-(acryloyloxy)ethyl] isocyanurate:

Poly-L-Tyrosine:

or
Dopamine:

12 . The substrate according to any preceding claim, wherein the monomer of formula (Ib) comprises any of the monomers selected from N-(4-Hydroxyphenyl)methacrylamide; 2-Norbornyl methacrylate; N-Phenylmethacrylamide; Cyclohexyl methacrylate; or Tris[2-(acryloyloxy)ethyl] isocyanurate; or
wherein the monomer of formula (Ib) comprises Poly-L-Tyrosine; or wherein the monomer of formula (Ib) comprises Dopamine.

13 . The substrate according to any preceding claim, wherein the substrate comprises a polymer blend of two or more different polymers.

14 . The substrate according to claim 13 , wherein the second and/or third polymers comprise a homopolymer formed from a monomer of formula (Ia) or (Ib); or a copolymer formed from one or more monomers of formula (Ia) and/or I(b); or a copolymer formed from one or more monomers of formula (Ia) and/or (Ib) and comprising HEMA.

15 . The substrate according to any preceding claim, wherein the polymer comprises a copolymer, and the copolymer comprises at least 10% HEMA.

16 . The substrate according to any preceding claim, wherein the copolymer comprises N-(4-Hydroxyphenyl)methacrylamide and HEMA.

17 . The substrate according to any preceding claim, wherein distinct regions of polymers comprise arrangements of strips, spots, lattices, or layers of one type of polymer alongside distinct regions or layers of another polymer.

18 . The substrate according to any preceding claim, wherein the substrate for cell culture is a coating on the surface of a cell culture device.

19 . The substrate according to any of claims 1 to 17 , wherein the substrate is in a solution, arranged to be applied to a surface of a cell culture device.

20 . The substrate according to any of claims 1 to 17 , wherein the substrate is in powdered form, wherein the substrate is arranged to be dissolved in solution and applied to a cell culture device.

21 . The substrate according to any preceding claim, wherein the cell culture device comprises a tissue culture plate.

22 . The substrate according to any preceding claim, wherein the cells to be cultured with or on the substrate comprise or consist of stem cells; partially or fully differentiated cells, such as fibroblasts; or progenitor cells; or combinations thereof.

23 . The substrate according to any preceding claim, wherein the cells comprise or consist of pluripotent stem cells or multipotent stem cells.

24 . The substrate according to any preceding claim, wherein the cells comprise or consist of human pluripotent stem cells (hPSCs) or human mesenchymal stem cells (hMSCs).

25 . A cell culture device comprising the substrate for culturing cells in accordance with any preceding claim.

26 . A polymer for use as a substrate for facilitating cell adhesion on a cell culture plate; wherein the polymer comprises
a homopolymer formed from a monomer of formula (Ia) or (Ib); a copolymer formed from one or more monomers of formula (Ia) and/or (Ib); or a copolymer formed from one or more monomers of formula (Ia) and/or I(b), and

Assignees

• Univ Nottingham

Inventors

• Alexander Morgan Russell
• Denning Chris
• Young Lorraine
• Celiz Adam
• Smith James
• Davies Martyn Christopher
• Patel Asha

Classifications

• C12N2523/00

  Culture process characterised by temperature · CPC title

• C12N2533/30

  Synthetic polymers · CPC title

• C08L2203/02

  for biomedical use · CPC title

• C12M47/02

  Separating microorganisms from the culture medium; Concentration of biomass (separating microorganisms from their culture media C12N1/02) · CPC title

• C08F20/18

  with acrylic or methacrylic acids · CPC title