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Method of constructing corneal endothelium of tissue engineering

A corneal endothelium and tissue engineering technology, applied in the field of tissue engineering corneal endothelium, can solve the problems of low corneal endothelium feasibility, poor corneal endothelial proliferation ability, and scarce donors, so as to improve corneal edema, facilitate transplantation, and reduce rejection. Effect

Active Publication Date: 2014-04-16
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The human corneal endothelium itself has poor proliferative ability and donors are scarce, so the feasibility of using human corneal endothelial cells to construct tissue-engineered corneal endothelium is low

Method used

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  • Method of constructing corneal endothelium of tissue engineering
  • Method of constructing corneal endothelium of tissue engineering

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Separation and acquisition of live amniotic membrane: the placenta was obtained from healthy cesarean section women (negative for HIV-I, hepatitis B, hepatitis C, and syphilis) who had signed an informed consent form in the hospital; The amniotic membrane was cut from the fresh placenta, the chorion was torn off, and then rinsed with 1×HBSS until smooth and transparent to obtain the live amniotic membrane, which was placed in SHEM medium. The whole process should be handled carefully and the epithelium must not be scratched.

[0034] (2) Spread the live cell-containing amniotic epithelium obtained in step (1) facing down on the culture insert, and put it into a six-well culture plate; add 200 μl of 2 mg / ml type IV collagenase to each of the above culture inserts , add 1.5ml SHEM medium to the culture plate below the plug-in to protect the epithelium, digest live amnion matrix at 37°C for 1h15min, after digestion, rinse with 1×PBS repeatedly to wash away type IV coll...

Embodiment 2

[0041] (1) Separation and acquisition of live amniotic membrane: the placenta was obtained from healthy cesarean section women (negative for HIV-I, hepatitis B, hepatitis C, and syphilis) who had signed an informed consent form in the hospital; The amniotic membrane was cut from the fresh placenta, the chorion was torn off, and then rinsed with 1×HBSS until smooth and transparent to obtain the live amniotic membrane, which was placed in SHEM medium. The whole process should be handled carefully and the epithelium must not be scratched.

[0042] (2) Spread the live cell-containing amniotic epithelium obtained in step (1) face down on the culture insert, and put it into a six-well culture plate; add 2mg / ml dispase II 200μl to each of the above culture inserts , add 1.5ml SHEM medium to the culture plate below the insert to protect the epithelium, digest live amnion matrix and basement membrane at 4°C for 2h30min, after digestion, rinse repeatedly with 1×PBS to wash away Dispase ...

Embodiment 3

[0049](1) Separation and acquisition of live amniotic membrane: the placenta was obtained from healthy cesarean section women (negative for HIV-I, hepatitis B, hepatitis C, and syphilis) who had signed an informed consent form in the hospital; The amniotic membrane was cut from the fresh placenta, the chorion was torn off, and then rinsed with 1×HBSS until smooth and transparent to obtain the live amniotic membrane, which was placed in SHEM medium. The whole process should be handled carefully and the epithelium must not be scratched.

[0050] (2) Spread the live cell-containing amniotic epithelium obtained in step (1) face down on the culture insert, and put it into a six-well culture plate; add 2mg / ml dispase II 200μl to each of the above culture inserts , add 1.5ml SHEM medium to the culture plate below the insert to protect the epithelium, digest live amnion matrix and basement membrane at 4°C for 2h30min, after digestion, rinse repeatedly with 1×PBS to wash away Dispase I...

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Abstract

The invention discloses a method of constructing a corneal endothelium of tissue engineering by utilizing a live amnion upper skin graft. The method comprises the steps: acquiring the live amnion upper skin graft; and constructing the corneal endothelium of the tissue engineering by virtue of the live amnion upper skin graft. With the adoption of the method, as shapes, the arrangement, tissue structures, biological functions and the like of cells of the adopted live amnion upper skin graft are similar to the corneal endothelium, the live amnion upper skin graft can be adopted to replace the corneal endothelium with dysfunction, the normal functions of cornea is maintained, and the cornea transparency is improved.

Description

technical field [0001] The invention belongs to the field of tissue engineering materials, in particular to a method for constructing tissue engineering corneal endothelium Background technique [0002] The amniotic membrane is the innermost membrane of the human placenta, which has the advantages of abundant sources, high transparency, good permeability, and does not contain HLA-A, B, C and DR antigen components; the organizational structure of the amniotic membrane includes a single layer of amniotic epithelial cells , basement membrane layer, and stroma layer. The amniotic epithelial cells are regular in shape and tightly arranged, and have good material exchange function. A mature amniotic membrane covers an area of ​​0.2 square meters and gathers 300 million amniotic epithelial cells. [0003] The corneal endothelium is formed by a layer of hexagonal endothelial cells, about 5 microns thick and 18-20 microns wide. It is in direct contact with the aqueous humor. This l...

Claims

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Application Information

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IPC IPC(8): C12N5/071A61L27/38
Inventor 李炜张丽颖邹笃雷刘祖国
Owner XIAMEN UNIV
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