Method for producing artificial skin model, and artificial skin model

Patent metadata

Field	Value
Publication number	US-10073085-B2
Application number	US-201214008348-A
Country	US
Kind code	B2
Filing date	Mar 29, 2012
Priority date	Mar 29, 2011
Publication date	Sep 11, 2018
Grant date	Sep 11, 2018

Abstract

Provided is a novel method capable of producing an artificial skin model. A method for producing an artificial skin model includes: providing coated cells (1), each of which is obtained by covering the surface of a cell (3) with a coating film (2) containing an extracellular matrix component; forming a dermis tissue layer (7), in which the coated cells (1) are laminated, by culturing the coated cells (1); and forming an epidermis layer (12) by arranging epidermis cells (8) on the dermis tissue layer (7).

First claim

The invention claimed is:

1. A production method for producing an artificial skin model, comprising:
providing cells;
providing, separately from the cells, a first material having an RGD sequence;
providing, separately from the cells, a second material which interacts with the first material;
forming extracellular matrix coated cells;
forming a dermis tissue layer, in which the extracellular matrix coated cells are laminated, by culturing the extracellular matrix coated cells; and
forming an epidermis layer by arranging epidermis cells on the dermis tissue layer,
wherein each of the extracellular matrix coated cells comprises:
a cell, and
a coating film coating the cell, wherein the coating film includes the first material and the second material, and at least one of the first material and the second material includes an extracellular matrix component,

the forming of the extracellular matrix coated cells comprises:
forming first material coated cells by contacting the first material to the cells by at least one selected from:
(a) soaking the cells in a liquid containing the first material,
(b) dripping the liquid containing the first material to the cells, and
(c) spraying the liquid containing the first material to the cells; and
then contacting the second material to the first material coated cells by at least one selected from:
(d) soaking the first material coated cells in a liquid containing the second material,
(e) dripping the liquid containing the second material to the first material coated cells, and
(f) spraying the liquid containing the second material to the first material coated cells,

wherein the first material is fibronectin, and the second material is gelatin.

2. The production method according to claim 1 , comprising forming a tissue ancillary organ in the dermis tissue layer.

3. The production method according to claim 1 , wherein the forming a dermis tissue layer comprises: forming a cell layer, in which the coated cells are laminated, by culturing the coated cells; arranging cells forming the tissue ancillary organ on the cell layer; and forming a cell layer in which the coated cells are laminated by arranging the coated cells on the cell layer in which the cells forming the tissue ancillary organ are arranged and culturing the coated cells arranged on the cell layer.

4. The production method according to claim 1 , wherein the forming a dermis tissue layer comprises arranging the coated cells so that the coated cells are laminated in at least two layers.

5. A production method for producing an artificial skin model, comprising:
providing cells;
providing, separately from the cells, a first material having an RGD sequence;
providing, separately from the cells, a second material which interacts with the first material;

forming extracellular matrix coated cells;
forming a dermis tissue layer, in which the extracellular matrix coated cells are laminated, by culturing the extracellular matrix coated cells; and
forming an epidermis layer by arranging epidermis cells on the dermis tissue layer,
wherein each of the extracellular matrix coated cells comprises:
a cell, and
a coating film coating the cell, wherein the coating film includes the first material and the second material, and at least one of the first material and the second material includes an extracellular matrix component,

the forming of the extracellular matrix coated cells comprises:
forming second material coated cells by contacting the second material to the cells by at least one selected from:
(a) soaking the cells in a liquid containing the second material,
(b) dripping the liquid containing the second material to the cells, and
(c) spraying the liquid containing the second material to the cells; and
then contacting the first material to the second material coated cells by at least one selected from:
(d) soaking the second material coated cells in a liquid containing the first material,
(e) dripping the liquid containing the first material to the second material coated cells, and
(f) spraying the liquid containing the first material to the second material coated cells,

wherein the first material is fibronectin, and the second material is gelatin.

6. The production method according to claim 5 , comprising forming a tissue ancillary organ in the dermis tissue layer.

7. The production method according to claim 5 , wherein the forming a dermis tissue layer comprises: forming a cell layer, in which the coated cells are laminated, by culturing the coated cells; arranging cells forming the tissue ancillary organ on the cell layer; and forming a cell layer in which the coated cells are laminated by arranging the coated cells on the cell layer in which the cells forming the tissue ancillary organ are arranged and culturing the coated cells arranged on the cell layer.

8. The production method according to claim 5 , wherein the forming a dermis tissue layer comprises arranging the coated cells so that the coated cells are laminated in at least two layers.

Assignees

• Akashi Mitsuru
• Matsusaki Michiya
• Sakura Takeshi
• Hashimoto Koji
• Shirakata Yuji
• Hirakawa Satoshi
• Univ Osaka
• Toray Industries

Inventors

• Akashi Mitsuru
• Matsusaki Michiya
• Sakura Takeshi
• Hashimoto Koji
• Shirakata Yuji
• Hirakawa Satoshi

Classifications

• C12N2502/1323

  Adult fibroblasts · CPC title

• G01N33/5088 Primary

  of vertebrates · CPC title

• C12N2533/52

  Fibronectin; Laminin · CPC title

• C12N2502/28

  Vascular endothelial cells · CPC title

• C12N2502/094

  keratinocytes · CPC title