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Method for promoting function and characteristic of corneal endothelial cells

A corneal endothelium and cell technology, which is applied in the field of promoting the function and characteristics of corneal endothelial cells, can solve the problem of inability to apply clinical treatment and related research, unable to maintain cell shape and function, and inability of human corneal endothelial cells to pass multiple times. Cell morphology and function, etc. problems, to achieve the effect of reliable and sufficient cell source, promotion of proliferation and repair ability, strong adherence and proliferation ability

Active Publication Date: 2017-04-26
吴欣怡
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Problems solved by technology

[0004] However, the in vitro culture of corneal endothelial cells has the following disadvantages: it cannot maintain its normal cell morphology and function after multiple passages, and it cannot be applied to clinical treatment and related research.
Therefore, there is an urgent need for a suitable culture method to solve the problem that human corneal endothelial cells cannot be passaged many times and maintain cell morphology and function.

Method used

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  • Method for promoting function and characteristic of corneal endothelial cells
  • Method for promoting function and characteristic of corneal endothelial cells
  • Method for promoting function and characteristic of corneal endothelial cells

Examples

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Effect test

Embodiment 1

[0056] 1. Isolate and culture human orbital adipose-derived mesenchymal stem cells and extract conditioned medium:

[0057] Isolation and culture of O-ASC: fat was obtained from patients undergoing blepharoplasty in the Medical Aesthetics Department of Qilu Hospital. Human orbital adipose tissue was collected under aseptic conditions. The following procedures were performed under aseptic conditions, rinsed 3 times with PBS, soaked in 75% ethanol for 30 s, rinsed 3 times with PBS, removed the blood vessels and connective tissue visible to the naked eye, and cut into pieces. to about 1mm 3 The particles were added with 2 volumes of 0.1% collagenase type I digestion solution and placed in a constant temperature shaker at 37°C for 1 h of slow shaking digestion. Add the same volume of low-glucose DMEM medium containing 10% FBS to neutralize, and centrifuge at 300 × g for 10 min. Discard the upper layer of fat and liquid, resuspend in sterile PBS, centrifuge at 300 × g for 5 min, ...

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Abstract

The invention discloses a method for promoting the function and characteristic of corneal endothelial cells. The method includes the following steps that human orbit adipose-derived stem cells are separated and cultured, and a condition culture medium is extracted; primary human corneal endothelial cells are separated and cultured; the condition culture medium is added into a basic culture medium for culturing the human corneal endothelial cells to culture and proliferate the human corneal endothelial cells. The wall adherence capacity and proliferation capacity of the human corneal endothelial cells cultured by the condition culture medium extracted from the human orbit adipose-derived stem cells are high. Through tests, by the utilization of the method, the human corneal endothelial cells cultured in an in-vitro mode can do down to future ten or more generations and are amplified by more times, and the shape and functions of the human corneal endothelial cells can be kept. Animal experiments verify that by the utilization of the method, the human corneal endothelial cells cultured in the in-vitro mode have an excellent cell repairing function.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering and ophthalmic repair, and in particular relates to a method for promoting the functions and properties of corneal endothelial cells. Background technique [0002] The cornea is divided into 5 layers, from front to back: epithelial layer, Descemet's layer, stroma, Descemet's layer and endothelial layer. Among them, the innermost endothelial cell layer is of great significance for maintaining corneal transparency and its normal physiological function. Endothelial cells are a single layer of cells with a height of about 50um and a width of about 20um. Human corneal endothelial cells (Hcecs) regulate corneal transparency through pump function and barrier function. With the increase of age, the density of corneal endothelial cells gradually decreases, and when the cell density is less than 500-800 cells / mm 2 Decompensation of the corneal endothelium will result, resulting in persistent c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079
CPCC12N5/0621C12N2500/14C12N2500/38C12N2500/84C12N2501/11C12N2501/90C12N2509/00C12N2502/1382C12N2501/113C12N2527/00C12N2320/32C12N5/0692C12N5/0653C12N2506/1384
Inventor 吴欣怡孙鹏
Owner 吴欣怡
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