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A method for constructing tissue-engineered corneal endothelium

A technology of corneal endothelial and tissue engineering, applied in artificial cell constructs, tissue regeneration, tissue culture, etc., can solve the problems of lack of sources of corneal endothelial cells and restrictions on the implementation of such operations in clinic, and achieve easy operation, wide distribution of sources, and preparation The effect of simple process

Active Publication Date: 2020-08-14
SHANDONG EYE INST
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Problems solved by technology

For this kind of disease, it can be treated by simply transplanting endothelium. At present, the commonly used methods include corneal endothelial transplantation with Descemet's membrane and endothelial cell transplantation with lamellar cornea, but the source of corneal donor or corneal endothelial cells The shortage seriously restricts the development of clinical such operations

Method used

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  • A method for constructing tissue-engineered corneal endothelium
  • A method for constructing tissue-engineered corneal endothelium

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Embodiment 1

[0029] 1. Preparation of Descemet's membrane of porcine cornea: take fresh and complete porcine cornea, soak and rinse with sterile saline containing blue-chain double antibody (green-chain double antibody (100×, purchased from Corning Company, USA)) for 20 minutes, and endothelial Put it face up under a stereomicroscope, use a cotton swab to carefully wipe off the porcine corneal endothelial cells, use toothed forceps to fix the porcine cornea, first tear off the residual iris root and expose the free part of the descemet layer, and then use flat-tipped forceps to carefully grasp the porcine cornea At the free part of the descemet's membrane, gently tear toward the center of the cornea to completely separate the descemet's membrane from the corneal stroma. Pay attention to the operating strength to avoid tearing the elastic layer.

[0030] Put the completely separated porcine corneal elastic membrane on a sterilized rubber ring with an inner diameter of 9.0mm, attach the endo...

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Abstract

The invention provides a method for establishing tissue engineering corneal endothelium. The method comprises the following steps: preparing a descemet membrane of swine corneal, culturing and inducing human multipotent stem cells, establishing tissue engineering corneal endothelium, and the like. Compared with an amniotic membrane or other carriers, the tissue engineering corneal endothelium established by using the method provided by the invention is not too soft and is good in transparency as the descemet membrane self is a normal cornea structure. Seed cells used in the method are induced and differentiated from the human multipotent stem cells, and are not limited by the multiplication capacity of primary cells. In addition, the tissue engineering corneal endothelium provided by the invention is simple in preparation process, wide in source and easy to operate.

Description

technical field [0001] The invention belongs to the field of tissue engineering, and in particular relates to a method for constructing tissue-engineered corneal endothelium. Background technique [0002] Corneal endothelium is a single layer of cells attached to Descemet's membrane and located on the innermost side of the cornea. It is in direct contact with aqueous humor and maintains corneal transparency and normal corneal thickness through its active fluid pump function. Adult corneal endothelial cells are unable to regenerate, and are mainly repaired by the expansion and migration of peripheral cells in the damaged area after injury. Various reasons such as Fuch corneal endothelial dystrophy, trauma and surgery can cause damage to corneal endothelial cells. When the cell density is lower than 500-1500 / mm 2 Decompensation of corneal endothelial function occurs immediately, leading to corneal edema and turbidity, showing bullous keratopathy. For this kind of disease, it...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/38C12N5/071
CPCA61L27/3633A61L27/3687A61L27/3691A61L27/3834A61L2430/16C12N5/0697C12N2506/02C12N2506/1346C12N2506/45C12N2533/90
Inventor 段豪云周庆军史伟云李文静
Owner SHANDONG EYE INST
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