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Tissue engineering skin construction method and tissue engineering skin produced by the same

A technology of tissue engineering skin and construction method, which is applied in the field of tissue engineering skin construction and tissue engineering skin prepared by using it, can solve the problems such as non-containment, achieve the effect of reducing variation, improving maturity and differentiation degree, and improving analysis accuracy

Inactive Publication Date: 2008-11-05
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 on the construction method of tissue engineering skin using fibroblasts to promote the differentiation of the stratum corneum, and the product does not contain fibroblasts.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] First, use Gibco's serum-free epidermal cell culture medium K-SFM as the base solution, and then add 35 ng of insulin, 200 μg of hydrocortisone, 30 mg of vitamin C, and 1.5 mM of calcium chloride according to the standard of 500 mL of base solution to prepare tissue engineering skin culture solution; Mix type I collagen, chitosan, hyaluronic acid and chondroitin sulfate according to the ratio of 10:2:1:1 mass parts, and then adjust the pH to 7.2 with 1M sodium hydroxide to prepare the gel biomaterial; Human-derived fibroblasts were treated with mitomycin C for 2.5 hours, and cultured with conventional cell culture medium DMEM with 10% serum to prepare human fibroblast feeder layer; the prepared human fibroblast feeder layer cells were divided into 5×10 5 / mL mixed well with 2mL of gel biomaterial to obtain a complex, and the complex was stored at 37°C, 5% CO 2 Cultivate in the incubator for 2h, then add 5×10 5 / mL epidermal cell suspension 1mL, continue to culture for ...

Embodiment 2

[0024] First, use Gibco's serum-free epidermal cell culture medium K-SFM as the base solution, then add 20 ng of insulin, 300 μg of hydrocortisone, 20 mg of vitamin C, and 2 mM of calcium chloride according to the standard of 500 mL of base solution to prepare tissue engineering skin culture solution; Type I collagen, chitosan, hyaluronic acid and chondroitin sulfate are mixed evenly according to the ratio of 12:2:1:1 by mass, and then the pH is adjusted to 7.2 with 1M sodium hydroxide to prepare the gel biomaterial; After the source fibroblasts were irradiated with 10Gy γ-rays for 18h, they were cultured in conventional cell culture medium DMEM with 2% serum to prepare human fibroblast feeder layer; the prepared human fibroblast feeder layer 5 / mL mixed well with 3mL of gel biomaterial to obtain a complex, and the complex was stored at 37°C, 5% CO 2 Cultivate in the incubator for 2h, then add 10×10 5 / mL epidermal cell suspension 1mL, continue to culture for 1h, add tissue e...

Embodiment 3

[0026] First, use Gibco's serum-free epidermal cell culture medium K-SFM as the base solution, then add 50 ng of insulin, 100 μg of hydrocortisone, 40 mg of vitamin C, and 1 mM of calcium chloride according to the standard of 500 mL of base solution to prepare tissue engineering skin culture solution; Type I collagen, chitosan, hyaluronic acid and chondroitin sulfate were mixed evenly according to the ratio of 9:3:1:1 by mass, and then the pH was adjusted to 7.3 with 1M sodium hydroxide to prepare the gel biomaterial; After the derived fibroblasts were treated with bleomycin for 5 hours, they were cultured with conventional cell culture medium DMEM with 8% serum to prepare human fibroblast feeder layer; 5 / mL mixed well with 3mL of gel biomaterial to obtain a complex, and the complex was stored at 37°C, 5% CO 2 Cultivate in the incubator for 2h, then add 1×10 5 / mL epidermal cell suspension 1mL, continue to culture for 1h, add tissue engineering skin culture medium 3mL. On t...

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PUM

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Abstract

The invention discloses a method for constructing tissue engineering skin, which comprises the steps as follows: (1) a tissue engineering skin nutrient solution is prepared; (2) a demic mechanocyte feeder layer is prepared; (3) a gelatin biomaterial is prepared; (4) the tissue engineering skin is constructed and cultivated. The method for constructing the tissue engineering skin adopts the adding of the mechanocytes after proliferation inhibition treatment to complete the interaction with epidermic cells so as to promote the mature and differentiation degree of the epidermic cells, has significant effect on improving the accuracy of compound toxicity detection in vitro to ensure that the prepared tissue engineering skin can more effectively resist the toxic action of the compound on competent cells and the forecast precision of the prepared tissue engineering skin to the compound toxicity is improved so as to create more effective animal substitution detection methods which are based on the tissue engineering skin.

Description

technical field [0001] The invention relates to a method for constructing skin using tissue engineering technology. 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. ...

Claims

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Application Information

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IPC IPC(8): A61L27/60G01N33/48
Inventor 邹运动董益阳王军兵郑洪艳程艳李铭杨晓冉
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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