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Method for constructing tissue engineering skin

A tissue-engineered skin and construction method technology, applied in the field of tissue-engineered skin construction, to achieve the effects of reducing variation, improving prediction sensitivity and accuracy, and facilitating quality control

Inactive Publication Date: 2009-07-22
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching, there is no technical literature about the method of using autocrine extracellular matrix to construct tissue-engineered skin.

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0016] A method for constructing tissue engineered skin utilizing autocrine extracellular matrix, comprising the following steps:

[0017] (1) Put the insert cell culture dish into the cell culture well plate, put 1×10 5 Fully mix human fibroblasts with 1mL cell culture medium to obtain a cell suspension, drop the cell suspension into an insert cell culture dish, and store at 37°C, 5% CO 2 Cultivate in the incubator for 10 days, change the medium twice a week, and obtain the complex of fibroblasts and autocrine extracellular matrix;

[0018] (2) Soak the complex of fibroblasts and autocrine extracellular matrix obtained in step (1) in sterile PBS at 37°C, 5% CO 2 Cultivate in an incubator for 48 hours to starve the fibroblasts to death, and obtain an autocrine extracellular matrix without living cells;

[0019] (3) UV irradiating the autocrine extracellular matrix obtained in step (2) without living cells for 1 h;

[0020] (4) Add 1×10 5 / mL human-derived epidermal cell su...

Embodiment 2

[0022] A method for constructing tissue engineered skin utilizing autocrine extracellular matrix, comprising the following steps:

[0023] (1) Put the insert cell culture dish into the cell culture well plate, put 5×10 5 Thoroughly mix human fibroblasts with 2mL cell culture medium to obtain a cell suspension, drop the cell suspension into an insert cell culture dish, and store at 37°C, 5% CO 2 Cultivate in the incubator for 20 days, change the medium twice a week, and obtain the complex of fibroblasts and autocrine extracellular matrix;

[0024] (2) Soak the complex of fibroblasts and autocrine extracellular matrix obtained in step (1) in sterile PBS at 37°C, 5% CO 2 Cultivate in an incubator for 48 hours to starve the fibroblasts to death, and obtain an autocrine extracellular matrix without living cells;

[0025] (3) UV irradiating the autocrine extracellular matrix obtained in step (2) without living cells for 3 hours;

[0026] (4) Add 5×10 5 / mL human-derived epiderma...

Embodiment 3

[0028] A method for constructing tissue engineered skin utilizing autocrine extracellular matrix, comprising the following steps:

[0029] (1) Put the insert cell culture dish into the cell culture well plate, put 10×10 5 Fully mix human fibroblasts with 3mL cell culture medium to obtain a cell suspension, drop the cell suspension into an insert cell culture dish, and store at 37°C, 5% CO 2 Cultivate in the incubator for 30 days, change the medium twice a week, and obtain the complex of fibroblasts and autocrine extracellular matrix;

[0030] (2) Soak the complex of fibroblasts and autocrine extracellular matrix obtained in step (1) in sterile PBS at 37°C, 5% CO 2 Cultivate in an incubator for 48 hours to starve the fibroblasts to death, and obtain an autocrine extracellular matrix without living cells;

[0031] (3) UV irradiating the autocrine extracellular matrix obtained in step (2) without living cells for 5 hours;

[0032] (4) Add 10×10 5 / mL human-derived epidermal c...

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Abstract

The invention relates to a method for constructing tissue-engineering skin. The construction method adopts an autocrine extracellular matrix of fibroblast as a biological scaffold of the tissue-engineering skin. The method utilizes the extracellular matrix excreted by the cultured fibroblast as the biological scaffold and human epidermal cells as seed cells, and the cultured tissue-engineering skin is closer to the structure of natural skin. The method has the advantages that: firstly, the method is simple and convenient, and favorable for synthesis and mass production of products because thematerials take the form of liquid; secondly, the obtained product has the structure which is closer to the natural skin, and the method is favorable for improving the sensibility and the precision ofprediction of the toxicity of compounds; thirdly, the product does not contain additional biomaterials, so that the risk of cross infection of zoonosis cannot occur; and fourthly, the obtained product is convenient for quality control, so that the variation between batches is reduced.

Description

technical field [0001] The invention relates to a method for constructing tissue engineered skin. Background technique [0002] The skin is the largest organ of the human body, and it consists of three parts, namely the epidermis, dermis and subcutaneous layer. The epidermis is located in the outermost layer of the skin, with a thickness between 0.06-0.8mm, and its microstructure is composed of basal layer, spinous cell layer, granular cell layer and stratum corneum. The stratum corneum is the final product of epidermal cell differentiation, 75%-80% of its composition is protein, 5%-15% is lipid, and a structure with a thickness of about 10 μm is composed of 10-15 layers of cells. The stratum corneum has many important physiological functions, which can effectively prevent the damage of external toxic compounds to the active cells of the skin, especially it has an extremely important barrier effect on the potential cytotoxicity of cosmetic ingredients. [0003] Tissue-engi...

Claims

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Application Information

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IPC IPC(8): A61L27/60A61L27/36A61F2/10
Inventor 邹运动董益阳程艳王军兵郑洪艳杨晓冉
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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