Cell culture substrate
The present invention is to provide a cell culture substrate including a block polymer including a segment having a lower critical solution temperature and a hydrophobic segment, the cell culture substrate further including an adhesive matrix, in which the adhesive matrix is an extracellular matrix and/or an adhesive synthetic matrix. Furthermore, the invention is to provide a cell culture substrate in which the extracellular matrix is at least one selected from laminin, fibronectin, vitronectin, cadherin, and fragments thereof, and/or the adhesive synthetic matrix is poly[2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl) ammonium hydroxide] or an oligopeptide-carrying polymer.
1. A cell culture substrate comprising a block polymer comprising a segment having a lower critical solution temperature and a hydrophobic segment, the cell culture substrate further including an adhesive matrix,
wherein a degree of polymerization of the segment having a lower critical solution temperature is 400 to 10,000, and
wherein the adhesive matrix is an extracellular matrix and/or an adhesive synthetic matrix.
2. The cell culture substrate according to claim 1 , wherein the extracellular matrix is at least one selected from laminin, fibronectin, vitronectin, cadherin, and fragments thereof.
3. The cell culture substrate according to claim 1 , wherein the adhesive synthetic matrix is poly[2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl) ammonium hydroxide] or an oligopeptide-carrying polymer.
4. The cell culture substrate according to claim 1 , wherein the hydrophobic segment is obtainable by polymerizing a monomer represented by the following Formula (1):
wherein in Formula (1), R 1 represents a hydrogen atom or a methyl group; and R 2 represents any one of a phenyl group, a carboxyalkyl group having an alkyl with 1 to 8 carbon atoms, a carboxyaralkyl group having an aralkyl with 7 or 8 carbon atoms, a group represented by the following Formula (2), or a group represented by the following Formula (3):
wherein in Formula (2), n represents 2 or 3; and R 3 represents an alkyl group having 1 to 3 carbon atoms,
wherein in Formula (3), R 4 and R 5 each independently represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms; and the total number of carbon atoms of R 4 and R 5 is 5 or more.
5. The cell culture substrate according to claim 1 , further comprising at least one protein selected from gelatin, collagen, and/or albumin on the cell culture substrate.
6. The cell culture substrate according to claim 5 , wherein the block polymer, at least one protein selected from gelatin, collagen, and/or albumin, and the adhesive matrix are sequentially laminated.
7. The cell culture substrate according to claim 1 , which is a dry cell culture substrate.
8. The cell culture substrate according to claim 1 , which is laminated on a supporting medium.
9. The cell culture substrate according to claim 8 , which has an average thickness of 1,000 nm or less.
10. A method for producing a dry cell culture substrate, the method comprising:
a step of coating a solution including at least one protein selected from gelatin, collagen, and/or albumin on the block polymer according to claim 1 ;
a step of further coating a solution including the adhesive matrix according to claim 1 thereon to obtain a cell culture substrate; and
a step of drying the obtained cell culture substrate.
11. A cell culture equipment comprising a supporting medium and the cell culture substrate according to claim 1 .
12. The cell culture substrate according to claim 2 , wherein the adhesive synthetic matrix is poly[2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl) ammonium hydroxide] or an oligopeptide-carrying polymer.
13. The cell culture substrate according to claim 2 , wherein the hydrophobic segment is obtainable by polymerizing a monomer represented by the following Formula (1):
wherein in Formula (1), R 1 represents a hydrogen atom or a methyl group; and R 2 represents any one of a phenyl group, a carboxyalkyl group having an alkyl with 1 to 8 carbon atoms, a carboxyaralkyl group having an aralkyl with 7 or 8 carbon atoms, a group represented by the following Formula (2), or a group represented by the following Formula (3):
wherein in Formula (2), n represents 2 or 3; and R 3 represents an alkyl group having 1 to 3 carbon atoms,
wherein in Formula (3), R 4 and R 5 each independently represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms; and the total number of carbon atoms of R 4 and R 5 is 5 or more.
14. The cell culture substrate according to claim 2 , further comprising at least one protein selected from gelatin, collagen, and/or albumin on the cell culture substrate.
15. The cell culture substrate according to claim 2 , which is a dry cell culture substrate.
16. The cell culture substrate according to claim 2 , which is laminated on a supporting medium.
17. A cell culture equipment comprising a supporting medium and the cell culture substrate according to claim 2 .
18. The cell culture substrate according to claim 3 , wherein the hydrophobic segment is obtainable by polymerizing a monomer represented by the following Formula (1):
wherein in Formula (1), R 1 represents a hydrogen atom or a methyl group; and R 2 represents any one of a phenyl group, a carboxyalkyl group having an alkyl with 1 to 8 carbon atoms, a carboxyaralkyl group having an aralkyl with 7 or 8 carbon atoms, a group represented by the following Formula (2), or a group represented by the following Formula (3):
wherein in Formula (2), n represents 2 or 3; and R 3 represents an alkyl group having 1 to 3 carbon atoms,
wherein in Formula (3), R 4 and R 5 each independently represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms; and the total number of carbon atoms of R 4 and R 5 is 5 or more.
19. The cell culture substrate according to claim 3 , further comprising at least one protein selected from gelatin, collagen, and/or albumin on the cell culture substrate.
20. The cell culture substrate according to claim 3 , which is a dry cell culture substrate.