Method for removing cells

Patent metadata

Field	Value
Publication number	US-9434936-B2
Application number	US-201113638795-A
Country	US
Kind code	B2
Filing date	Mar 28, 2011
Priority date	Apr 2, 2010
Publication date	Sep 6, 2016
Grant date	Sep 6, 2016

Abstract

A method for separating cells which includes: adhering cells to the surface of a cell culture substrate containing a photo-acid generator that generates an acidic substance upon irradiation with active energy rays, and irradiating only a partial region of the cell culture substrate with the active energy rays to selectively remove the cells within the partial region, thereby separating the cells within the partial region and cells in other regions.

First claim

The invention claimed is:

1. A method for removing target cells from a cell culture substrate, the method comprising:
providing a cell culture substrate including a substrate and a layer containing a photo-acid generator formed on a surface of the substrate:
adhering cells to the surface of the layer containing the photo-acid generator which generates an acidic substance upon irradiation with light,
generating the acidic substance by irradiating only a region of the substrate containing the photo-acid generator layer beneath target cells to be removed, and
removing the target cells from the region of the irradiated substrate by the action of the acidic substance generated from the irradiated layer containing the photo-acid generator beneath the target cells.

2. The method for removing target cells according to claim 1 , wherein the removal is conducted by killing the target cells by the action of the acidic substance generated from the irradiated layer containing the photo-add generator beneath the removal target cells.

3. The method for removing cells according to claim 1 , wherein the substrate comprises a polyvinylpyridine-based resin.

4. The method for removing target cells according to claim 1 , wherein
a layer containing an adhesion inhibitor is formed on only a partial region of the surface of the layer containing the photo-acid generator, and
the layer containing the photo-acid generator, which has the target cells adhered thereto in a region in which the layer containing an adhesion inhibitor is not formed, is irradiated with light.

5. The method for removing cells according to claim 1 , wherein prior to irradiation with the light, a portion of the cells are labeled, and a region to be irradiated with light is determined based on positioning information for the labeled cells.

6. The method for removing cells according to claim 1 , wherein
the photo-acid generator is at least one member selected from the group consisting of a naphthaleneimide-based sulfonic acid derivative, a thioxanthone-based sulfonic acid derivative, 1,8-naphthalenedicarboxylic acid imidomethylsulfonate, 1,8-naphthalenedicarboxylic acid imidotosylsulfonate, and sulfonium salts or iodonium salts having an anion which is tetrafluoroborate (BF 4 − ) or hexafluorophosphate (PF 6 − ).

7. The method for removing cells according to claim 1 , wherein
the photo-acid generator is at least one member selected from the group consisting of a compound represented by formula (1) and a compound represented by formula (2) shown below:

8. The method for removing cells according to claim 1 , wherein
the photo-acid generator includes a compound which generates an acidic substance by irradiation with the light, and
the acidic substance is a polymeric compound.

9. The method for removing cells according to claim 8 , wherein the photo-acid generator is at least one member select from the group consisting of a compound represented by formula (3a), a compound represented by formula (3b), a compound represented by formula (3c) and a compound represent by formula (3d):

wherein
for the compound represented by formula 3(a): R 1 is Me and R 2 is OMe;
for the compound represented by 3(b): R 1 is H and R 2 is OMe;
for the compound represented by 3(c): R 1 is H and R 2 is NH-iPr; and
for the compound represented by 3(d): R 1 is H and R 2 is NH-iPr; wherein x and y are integers of 1 or greater.

10. The method for removing cells according to claim 1 , wherein the light has a wavelength of 200 to 600 nm.

11. The method for removing cells according to claim 1 , wherein a light source of the light is an ultraviolet lamp or a visible light lamp.

12. The method for removing target cells according to claim 1 , wherein the step of removing the target cells is by washing the irradiated substrate with a culture medium or a buffer solution thereby detaching the target cells.

Assignees

• Sumaru Kimio
• Takagi Toshiyuki
• Kikuchi Kyoko
• Sato Taku
• Yamaguchi Manae
• Kanamori Toshiyuki
• Nat Inst Advanced Ind Science & Tech

Inventors

• Sumaru Kimio
• Takagi Toshiyuki
• Kikuchi Kyoko
• Sato Taku
• Yamaguchi Manae
• Kanamori Toshiyuki

Classifications

• C12M23/20

  Material Coatings (immunocoatings C12M25/00) · CPC title

• C12M25/06

  Plates; Walls; Drawers; Multilayer plates · CPC title

• C12M33/00

  Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus (chemical or physical laboratory apparatus in general B01L, devices for taking cell samples A61B10/0045, withdrawing or distributing predetermined quantities of fluid B01L99/00) · CPC title

• C12N1/02

  Separating microorganisms from their culture media · CPC title

• C12N13/00 Primary

  Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title