Cell culture medium and immobilized preparation of cell adhesion protein or peptide

• US 8,304,238 B2
• Filed: 03/24/2004
• Issued: 11/06/2012
• Est. Priority Date: 03/24/2003
• Status: Expired due to Fees

• Current Assignee: Katsumi Mochitate
• Original Assignee: Katsumi Mochitate, Nat'L Institute For Environmental Studies
• Inventors: Mochitate, Katsumi
• Primary Examiner(s): HANLEY, SUSAN MARIE
• Application Number:
  US10/551,052
• Publication Number:
  US 20060263878A1
• Time in Patent Office: 3,149 Days
• Field of Search: 435/395, 435/396, 435/402, 435/180, 530/815
• US Class Current: 435/402
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• 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

Abstract

The invention relates to an immobilized preparation of a cell adhesion protein or peptide for cell culture having a hydrophobic cell culture substrate, a cell adhesion protein or peptide and a hydrophobic binding-adsorptive polymer to which the cell adhesion protein or peptide has been covalently bound. The hydrophobic binding-adsorptive polymer to which the cell adhesion protein or peptide is covalently bound is adsorbed to the hydrophobic cell culture substrate by hydrophobic binding and not by chemical bonding. The hydrophobic binding-adsorptive polymer is a copolymer of maleic anhydride and styrene, a copolymer of maleic anhydride and butyl vinyl ether or a copolymer of maleic anhydride and hexyl vinyl ether.

Claims

1. An immobilized preparation of a cell adhesion protein or peptide for cell culture, comprising:

a hydrophobic cell culture substrate,

a cell adhesion protein or peptide,

a hydrophobic binding-adsorptive polymer to which the cell adhesion protein or peptide has been covalently bound,

wherein the hydrophobic binding-adsorptive polymer to which the cell adhesion protein or peptide is covalently bound, is adsorbed to the hydrophobic cell culture substrate by hydrophobic binding and not by chemical bonding, and

wherein the hydrophobic binding-adsorptive polymer is selected from a copolymer of maleic anhydride and styrene;

a copolymer of maleic anhydride and butyl vinyl ether; and

a copolymer of maleic anhydride and hexyl vinyl ether.

2. A method for producing an immobilized preparation of a cell adhesion protein or peptide for cell culture, comprising:

(a) reacting a functional group of a hydrophobic binding-adsorptive polymer with a cell adhesion protein or peptide, the functional group being capable of reacting with the cell adhesion protein or peptide to produce a hydrophobic binding-adsorptive polymer covalently bound to the cell adhesion protein or peptide, and

(b) coating a hydrophobic cell culture substrate with the hydrophobic binding-adsorptive polymer covalently bound to the cell adhesion protein or peptide from step (a), so that the hydrophobic binding-adsorptive polymer covalently bound to the cell adhesion protein or peptide adsorbs to the hydrophobic cell culture substrate by hydrophobic binding and not by chemical bonding,

wherein the hydrophobic binding-adsorptive polymer is selected from a copolymer of maleic anhydride and styrene;

a copolymer of maleic anhydride and butyl vinyl ether; and

a copolymer of maleic anhydride and hexyl vinyl ether.

3. A method for producing an immobilized preparation of a cell adhesion protein or peptide for cell culture, comprising:

(a) coating a hydrophobic cell culture substrate with a hydrophobic binding-adsorptive polymer, so that hydrophobic binding-adsorptive polymer adsorbs to the hydrophobic cell culture substrate by hydrophobic binding and not by chemical bonding to produce a hydrophobic cell culture substrate having the hydrophobic binding-adsorptive polymer adsorbed thereto;

(b) reacting a functional group of the hydrophobic binding-adsorptive polymer bound to the hydrophobic cell culture substrate from step (a) with a cell adhesion protein or peptide, the functional group being capable of reacting with the cell adhesion protein or the peptide to produce a hydrophobic cell culture substrate having a hydrophobic binding-adsorptive polymer adsorbed thereto wherein the functional group of the hydrophobic binding-adsorptive polymer bound thereto is covalently bound to the cell adhesion protein or peptide, and

wherein the hydrophobic binding-adsorptive polymer is selected from a copolymer of maleic anhydride and styrene;

a copolymer of maleic anhydride and butyl vinyl ether; and

a copolymer of maleic anhydride and hexyl vinyl ether.