Corneal endothelium-like sheet and method of constructing the same

• US 20050214259A1
• Filed: 02/13/2003
• Published: 09/29/2005
• Est. Priority Date: 04/30/2002
• Status: Abandoned Application

• Current Assignee: ArBlast Co., Ltd.
• Original Assignee: ArBlast Co., Ltd.
• Inventors: Ishino, Yutaka, Sano, Yoichiro, Nakamura, Takahiro, Kinoshita, Shigeru
• Application Number:
  US10/510,958
• Publication Number:
  US 20050214259A1
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61L 2430/16 for reconstruction of eye parts, e.g. intraocular lens, cornea
• CPC Class Codes: A61L 27/3808 Endothelial cells
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2533/92 Amnion; Decellularised dermis or mucosa
• CPC Class Codes: C12N 5/0621 Eye cells, e.g. cornea, iris pigmented cells photoreceptors C12N5/062

Abstract

It is our intention to provide a transplantation material applicable to the treatment of various diseases that require corneal endothelium transplantation. Corneal endothelial cells are collected, proliferated and then sown on amnion and cultured. Thus, a corneal endothelium-like sheet, which has a cell layer made up of the corneal endothelium-origin cells formed on the amnion, can be obtained.

Claims

1. A corneal endothelium-like sheet, comprising:

a collagen layer; and

a cell layer formed on the collagen layer, the cell layer consisting of cells derived from corneal endothelium.

2. A corneal endothelium-like sheet according to claim 1, wherein the collagen layer is derived from amniotic membrane.

3. A corneal endothelium-like sheet according to claim 1, wherein the collagen layer consists of amniotic membrane from which the epithelium has been removed.

5. A corneal endothelium-like sheet according to claim 1, wherein the cell layer has a monolayer structure.

6. A corneal endothelium-like sheet according to claim 1, wherein the cell density of the cell layer is about 2000 cells/mm² to about 4000 cells/mm².

7. A corneal endothelium-like sheet according to claim 1, wherein the plane view shape of the cells derived from corneal endothelium is hexagonal.

8. A corneal endothelium-like sheet according to claim 1, wherein the cells derived from corneal endothelium are arranged regularly in the cell layer.

4. A corneal endothelium-like sheet comprising a cell layer consisting of cells derived from corneal endothelium.

9. A method for constructing a corneal endothelium-like sheet, the method comprising the following steps:

a) culturing and proliferating collected corneal endothelial cells;

b) collecting the proliferated corneal endothelial cells and producing a cell suspension; and

c) planting the cell suspension on a collagen layer and culturing thereof.

10. A method for producing a corneal endothelium-like sheet according to claim 9, wherein the following step is carried out after step b):

b-1) increasing the cell density in the cell suspension using centrifugation.

11. A method for producing a corneal endothelium-like sheet according to claim 9, wherein centrifugation is carried out after planting the cell suspension in step c).

12. A method for producing a corneal endothelium-like sheet according to claim 9, wherein step c) comprises the following steps:

c-1) placing a container in a culture container, the container having a bottom surface consisting of a membrane with a pore size capable of allowing a culture solution to pass through;

c-2) forming a collagen layer on the bottom face of the container;

c-3) planting the cell suspension on the collagen layer;

c-4) carrying out centrifugation;

c-5) culturing.

13. A method for producing a corneal endothelium-like sheet according to claim 9, wherein the collagen layer is derived from amniotic membrane.

14. A method for producing a corneal endothelium-like sheet according to claim 9, wherein the collagen layer consists of amniotic membrane from which the epithelium has been removed.