Corneal epithelial sheet, method of constructing the same and transplantation method using the sheet

A production method and corneal epithelial cell technology are applied in the application field of the film, which can solve the problems of decreased vision, limbal depletion, and inability to maintain the cornea for a long time, and achieve the effects of high viability and high safety.

Inactive Publication Date: 2007-03-14
阿如布勒斯特有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Then, due to the extinction of the stem cell tissue, the defect site is covered by the surrounding conjunctival epithelium, and the transparency is insufficient, causing an extreme decrease in vision
In this way, due to the depletion of the corneal limbus in the above-mentioned pathological conditions, the transplanted cornea cannot be maintained for a long time only by cornea transplantation.

Method used

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  • Corneal epithelial sheet, method of constructing the same and transplantation method using the sheet
  • Corneal epithelial sheet, method of constructing the same and transplantation method using the sheet

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Fabrication of Three-dimensional Corneal Epithelial Sheets Using Collagen Gels

[0036] 1. Modulation of Corneal Epithelial Cells

[0037] 1-1. Preparation of cornea

[0038] Cornea was a donor material purchased from Northwest Lions Eye Bank (Seattle, USA).

[0039] 1-2. Serum-free culture method of corneal epithelial cells

[0040] The cornea was transferred to a Petri dish added with Duchenne's phosphate buffered solution (PBS), and one side of the edge was cut to 2-3 mm with a scalpel under a solid microscope. The cut edge was washed several times with PBS, and sterilized by immersing in 70% ethanol for 1 minute. After washing with PBS, soak in a dispase solution (dispase II, contract alcohol company, 250 units / ml, Duchenne's modified MEM medium; DMEM), and let stand at 4° C. overnight (18 to 24 hours). The next day, under a stereomicroscope, peel the epithelium away from the parenchyma with forceps. The peeled corneal epithelium was washed with DMEM and then w...

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Abstract

A corneal epithelial sheet having potential for the achievement of a favorable therapeutic effect and being highly safe in transplantation. The corneal epithelial sheet is constructed by: (a) preparing corneal epithelial cells; (b) separately culturing human fibroblasts in a collagen gel; (c) sowing or placing the corneal epithelial cells on the collagen gel; and (d) culturing and proliferating the corneal epithelial cells in the absence of xenogeneic animal cells.

Description

technical field [0001] The present invention relates to corneal epithelial sheets. Specifically, the present invention relates to the production of a corneal epithelial sheet by culturing and proliferating corneal epithelial cells on a collagen gel and a method for producing the sheet without using cells derived from a different species of animal as feeder cells. And the application of the film (transplantation method, etc.). Background technique [0002] One of the tissues expected to be used in regenerative medicine is the cornea. The cornea, the outermost layer that makes up the optical system of the eye, is a transparent, avascular tissue that facilitates good vision by forming a smooth surface with tears. In addition, corneal and conjunctival epithelial cells are often in contact with the outside world, and have the function of protecting the eyeball from foreign substances such as microorganisms and light such as ultraviolet rays from the outside world. That is, cor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/00C12N5/06A61L27/24A61L27/38C12N5/071
CPCC12N5/0621A61L27/3895C12N2502/1323A61L27/24A61L27/3869A61L27/3813C12N2533/54A61L2430/16A61L27/3804C12N5/0602A61L27/00C12N11/16
Inventor 桥本公二白方裕司大桥裕一羽室淳尔
Owner 阿如布勒斯特有限公司
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