Cell treatment method, laser processing machine, and cell culture vessel

Patent metadata

Field	Value
Publication number	US-10876086-B2
Application number	US-201615576228-A
Country	US
Kind code	B2
Filing date	Mar 25, 2016
Priority date	Jun 1, 2015
Publication date	Dec 29, 2020
Grant date	Dec 29, 2020

Abstract

A method of killing specific cells from among a group of cells cultured in a culture vessel by quick and brief laser treatment, the cell culture vessel comprising a main body and a to-be-irradiated layer attached to the main body, the to-be-irradiated layer containing an ingredient capable of absorbing laser light upon laser irradiation, the group of cells being cultured on the surface of the to-be-irradiated layer, the method comprising:
applying laser light to a partial area of the to-be-irradiated layer directly below the specific cells.

First claim

The invention claimed is:

1. A method of killing specific cells from among a group of cells cultured in a vessel comprising a main body and a layer attached to the main body, wherein the layer contains an ingredient capable of generating heat upon laser irradiation and the group of cells are cultured on the top surface of the layer, the method comprising applying a laser light to a partial area of the bottom surface of the layer by moving the laser light along a plane parallel to the partial area, wherein:
the partial area of the layer irradiated with the laser light absorbs energy of the laser light and thereby generates heat that kills unwanted cells that are present directly above the partial area,
the laser light does not instantly kill the specific cells upon irradiation,
the laser light output is 5 W or less,
the laser light wavelength is 380 nm or greater,
the laser light moving rate is 2000 mm/sec or less, and
the thickness of the layer is 10 mm or less.

2. The method according to claim 1 , wherein the laser light is applied multiple.

3. The method according to claim 1 , further comprising dividing the group of cells into a plurality of portions,
wherein the laser light is applied to a partial area of the layer directly below a boundary between two of the portions.

4. The method according to claim 1 , wherein the layer further comprises a polymer and the ingredient comprises an organic compound derivative.

5. The method according to claim 4 , wherein the ingredient is selected from the group consisting of azobenzene, diarylethene, spiropyrane, spirooxazines, fulgides, leucopigments, indigo dye, carotinoids, flavonoids and quinoids.

6. The method according to claim 5 , wherein the polymer comprises poly(methyl methacrylate) and the ingredient comprises azobenzene.

7. The method according to claim 4 , wherein the polymer is selected from the group consisting of acrylic polymers, polystyrene polymers, polyolefin polymers, polyvinyl acetate, polyvinyl chloride, polycarbonate polymers, and epoxy polymers.

8. The method according to claim 1 , wherein the top surface of the layer containing the ingredient is coated with an extracellular matrix.

9. The method according to claim 1 , wherein the laser light is continuously moved along a plane which is parallel to a plane of the partial area.

10. The method according to claim 1 , wherein the rate of moving the laser light is based on the output of the laser light.

11. The method according to claim 1 , wherein the wavelength of the laser light is in a range of 380 nm to 808 nm.

12. The method according to claim 1 , wherein an energy amount of the laser light is 25.9 J/cm 2 or less.

13. The method according to claim 1 , wherein the laser light is applied by moving a processing nozzle including a lens for gathering the laser light prior to irradiation of the layer.

Assignees

• Kataoka Corp
• Aist

Inventors

• Suzuki Masami
• Nishi Norio
• Matsumoto Junichi
• Sumaru Kimio
• Kanamori Toshiyuki

Classifications

• C12M35/02 Primary

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title

• C12M31/02

  located outside the reactor · CPC title

• C12N1/00

  Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor · CPC title

• C12M25/06

  Plates; Walls; Drawers; Multilayer plates · CPC title

• C12M23/20

  Material Coatings (immunocoatings C12M25/00) · CPC title