Method for in-vitro construction of tissue-engineered corneal endothelium

A corneal endothelium and tissue engineering technology, used in tissue regeneration, tissue culture, artificial cell constructs, etc., can solve the problem that the cornea is not suitable for clinical treatment, and achieve the advantages of adhesion and in vitro culture observation, mild edema, and solve the source problem. Effect

Active Publication Date: 2017-07-11
SHANDONG EYE INST
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Problems solved by technology

[0003] The applicant prepared ultra-thin decellularized porcine corneal posterior stroma containing Descemet's lamina, combined with corneal endothelial cells derived from human pluripotent stem cells to construct tissue-engineered corneal endothelium in vitro, which can not only solve the problem of corneal donor sources, but also overcome tissue engineering. The difficult problem that the thickened cornea is not suitable for clinical treatment

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  • Method for in-vitro construction of tissue-engineered corneal endothelium
  • Method for in-vitro construction of tissue-engineered corneal endothelium

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Embodiment

[0039] 1. Preparation of ultra-thin decellularized porcine corneal posterior stroma: fresh porcine corneal posterior stroma with a diameter of 8 mm and a thickness of 0.09 mm was cut with a femtosecond laser, and the endothelial cell side was marked with Descemet's membrane preserved. The cut porcine corneal posterior stroma was sealed in a plastic bag filled with protective solution, and the protective solution was used to protect it throughout the process. Break the cells under the condition of 200MPa high static pressure, treat twice, each time for 2 minutes; take out the porcine corneal stroma, put it in the protective solution of 0.3% sodium lauryl sulfate and 200U / ml DNase, set the shaking The bed speed was 100 rpm, and the DNA components in the cornea were removed by digestion at 25°C for 3 hours. After completion, the posterior stroma of the porcine cornea was rinsed in the protective solution for 3 hours. After rinsing, the decellularized porcine corneal stromal endot...

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Abstract

The invention provides tissue-engineered corneal endothelium constructed in vitro and a preparation method thereof. The preparation method comprises the following steps: preparation of an ultrathin acellular porcine corneal posterior matrix; culture; induction of human multi-potent stem cells; construction of the tissue-engineered corneal endothelium; etc. According to the invention, the ultrathin acellular porcine corneal posterior matrix containing the Descemet's membrane is prepared and human multi-potent stem cell-derived corneal endothelial cells are cooperatively used for in-vitro construction of the tissue-engineered corneal endothelium; so problems in sources of cornea donors are overcome, and the problem that tissue-engineered corneas are too thick for clinical treatment is overcome.

Description

technical field [0001] The invention belongs to the field of tissue engineering, in particular to a method for constructing tissue-engineered corneal endothelium in vitro. Background technique [0002] The corneal endothelium is a hexagonal monolayer of cells attached to Descemet's membrane, which maintains corneal transparency and normal corneal thickness through its active fluid pump function. The proliferation ability of human corneal endothelial cells is extremely limited, and the damage is mainly repaired by the expansion and migration of peripheral cells in the damaged area. Various reasons such as Fuch corneal endothelial dystrophy, trauma, surgery or inflammation can cause damage to corneal endothelial cells. When the density of corneal endothelial cells is lower than 500-1500 / mm2, corneal endothelial function decompensation occurs, resulting in corneal edema Cloudy, bullous keratopathy, corneal endothelial transplantation with Descemet's membrane or endothelial cel...

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Application Information

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IPC IPC(8): A61L27/36A61L27/38C12N5/071
CPCA61L27/3633A61L27/3687A61L27/3691A61L27/3834A61L2430/16C12N5/0697C12N2506/02C12N2506/1346C12N2506/45C12N2533/90
Inventor 史伟云周庆军段豪云董沐晨
Owner SHANDONG EYE INST
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