# US 9,434,927 B2 · Method for manufacturing cell culture substrate

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- Number: US 9,434,927 B2
- Document ID: US9434927B2
- Status: Granted
- Kind: B2
- Application number: 13/820,037
- Filed: 2011-08-30
- Published: 2013-08-29
- Granted: 2016-09-06
- Earliest claimed: 2010-08-31
- Assignees: Osaka Organic Chemical Industry Ltd.
- Applicants: Hiromi Kitano; Yoshiyuki Saruwatari; Kazuyoshi Matsuoka
- Inventors: Hiromi Kitano; Yoshiyuki Saruwatari; Kazuyoshi Matsuoka
- Web page: https://patentgrep.com/patent/US9434927B2

## CPC

- C12N 5/0686: {Kidney cells}
- C12N 5/0068: {General culture methods using substrates (for specific animal cell type C12N5/06)}
- C12N 2533/30: Synthetic polymers
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

A method for manufacturing a cell culture substrate obtained by forming a coating layer of a polymer on the surface of a substrate, wherein the polymer is formed by polymerizing a monomer component containing a nitrogen atom-containing monomer represented by formula (I): wherein R1 is hydrogen atom or methyl group, R2 is an alkylene group having 1 to 6 carbon atoms, each of R3 and R4 is independently an alkyl group having 1 to 4 carbon atoms, R5 is an alkylene group having 1 to 4 carbon atoms, and Y is oxygen atom or —NH— group, includes irradiating the polymer with ion beam at a predetermined position of the coating layer of the polymer, to remove the coating layer of the polymer irradiated with the ion beam.

## Claims

### Claim 1

A method for manufacturing a cell culture substrate comprising a substrate and a coating layer of a polymer formed on the surface of the substrate, said method comprising the steps of,
polymerizing a monomer component comprising a nitrogen atom-containing monomer represented by the formula (I) on the surface of a substrate having hydroxyl groups thereon, to form a coating layer of the polymer on said surface:
wherein R1 is hydrogen atom or methyl group, R2 is an alkylene group having 1 to 6 carbon atoms, each of R3 and R4 is independently an alkyl group having 1 to 4 carbon atoms, R5 is an alkylene group having 1 to 4 carbon atoms, and Y is oxygen atom or —NH— group, and
irradiating the coating layer of the polymer at a predetermined position with ion beam, to remove the coating layer of the polymer irradiated with the ion beam at the predetermined position.

### Claim 2 (depends on 1)

The method for manufacturing a cell culture substrate according to claim 1, wherein the monomer component further comprises at least one copolymerizable monomer selected from the group consisting of an alkyl (meth)acrylamide-based monomer, a polyethylene glycol-based monomer, a polypropylene glycol-based monomer, a sugar backbone-containing monomer, a phosphobetaine-based monomer and a sulfobetaine-based monomer.

### Claim 3 (depends on 1)

A cell culture substrate obtained by the method for manufacturing a cell culture substrate according to claim 1.

### Claim 4 (depends on 1)

The method for manufacturing a cell culture substrate according to claim 1, wherein the substrate having hydroxyl groups is a glass substrate.

### Claim 5 (depends on 4)

The method for manufacturing a cell culture substrate according to claim 4, wherein the glass substrate is a glass plate.

### Claim 6 (depends on 1)

The method for manufacturing a cell culture substrate according to claim 1, wherein R2 is a methylene group.

### Claim 7 (depends on 1)

The method for manufacturing a cell culture substrate according to claim 1, wherein R2 is an ethylene group.

### Claim 8 (depends on 1)

The method for manufacturing a cell culture substrate according to claim 1, wherein R3 and R4 are methyl groups.

### Claim 9 (depends on 1)

The method for manufacturing a cell culture substrate according to claim 1, wherein R3 and R4 are ethyl groups.

### Claim 10 (depends on 1)

The method for manufacturing a cell culture substrate according to claim 1, wherein R5 is a methylene group.

### Claim 11 (depends on 1)

The method for manufacturing a cell culture substrate according to claim 1, wherein R5 is an ethylene group.