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Production method of human corneal epithelial sheet

A technology of corneal epithelial cells and a manufacturing method is applied in the field of human corneal epithelial sheets for transplantation, which can solve the problems of insufficient donors and low survival rate of corneal epithelium, and achieve the effect of solving the shortage of donors

Active Publication Date: 2018-09-21
JCR PHARMA +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Corneal epithelial transplantation has a low survival rate due to rejection in corneas that are inflamed due to corneal epithelial stem cell deficiency
[0006] In addition, corneal epithelial transplantation also has the problem of insufficient donors who cannot obtain enough corneal epithelium for transplantation

Method used

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  • Production method of human corneal epithelial sheet
  • Production method of human corneal epithelial sheet
  • Production method of human corneal epithelial sheet

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Embodiment

[0156] Hereinafter, the present invention will be described in further detail with reference to examples, but the present invention is not intended to be limited to the examples.

[0157] 〔Preparation of supporting cells〕

[0158] The cryopreserved human mesenchymal stem cells (hMSCs) were quickly thawed in a thermostat at 37° C., added with DMEM / 10% FBS medium and suspended, centrifuged, and the cells were pelleted and recovered. The recovered cells were suspended in DMEM / 10% FBS medium, and the number of living cells was measured. Inoculate on the bottom well (bottom well) of 6 wells of cell culture insert matching plate (6 well matching plate, BD Biosciences company), so that the density is 15000-25000 / cm 2 , in DMEM / 10% FBS medium, in 5% CO 2 In the presence, cultured at 37°C for 1-3 days. Before hMSCs were used as supporting cells for the primary culture (P0 culture) of human corneal epithelial cells, the DMEM / 10% FBS medium was removed from the bottom well, and 1.5 mL...

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Abstract

Subject: To provide a method for producing human corneal epithelial sheets for transplantation to patients with corneal diseases, and for culturing cells derived from human corneal epithelium obtained by culturing human corneal epithelial cells on stromal amniotic membrane. Solution: culture method of human corneal epithelial cells using human mesenchymal stem cells as supporting cells, and using various combinations of ROCK inhibitors, phosphodiesterase inhibitors, MAP kinase inhibitors, and TGF‑β receptor inhibitors culture medium of human corneal epithelial cells.

Description

technical field [0001] The present invention relates to a human corneal epithelial sheet for transplantation. Specifically, it relates to a method for producing a human corneal epithelial sheet using cells derived from human corneal epithelium obtained by culturing human corneal epithelial cells. Background technique [0002] The cornea is a transparent tissue, which is composed of 5 cell layers of corneal epithelium, Bowman's membrane, corneal stroma, Bowman's membrane, and corneal endothelium from the outside to the inside. It has no blood vessels but nerve distribution. The corneal epithelium is non-keratinized stratified squamous epithelium with a thickness of about 50 μm, which constitutes a part of the ocular surface, restricts the permeation of substances, and functions as a barrier to prevent bacteria, fungi, etc. from entering the cornea. The tight junctions present between the outermost cells play an important role in maintaining the barrier function of the corneal...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A61L27/00
CPCC12N5/0621C12N2501/15C12N2501/727C12N2501/999A61L27/3813A61L2430/16A61P27/02A61K35/36C12N5/0602C12N2502/1352
Inventor 今川究前田谦一细田勇喜横山修一奥村直毅小泉范子木下茂
Owner JCR PHARMA
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