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Preparation method of novel tissue engineering skin

A tissue-engineered skin, a new type of technology, applied in biochemical equipment and methods, tissue culture, epidermal cells/skin cells, etc., can solve the problem of immunogenicity of natural scaffolds that do not have biological activity, and achieve the effect of low immunogenicity

Inactive Publication Date: 2017-10-17
林翠霞
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the deficiencies of the prior art, the purpose of the present invention is to provide a method for using amniotic membrane decellularized scaffold and mesenchymal stem cells as a carrier to construct a keratinocyte-like in vivo culture environment, and adopt a serum-free culture system to cultivate and prepare a new tissue engineering method. Solve the problem that in the prior art, the artificial scaffold does not have the biological activity and the immunogenicity of the natural scaffold

Method used

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  • Preparation method of novel tissue engineering skin
  • Preparation method of novel tissue engineering skin
  • Preparation method of novel tissue engineering skin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 The present invention is realized through the following steps.

[0027] 1. Skin tissue preparation:

[0028] (1) Quickly place the excised autologous skin in sterile PBS solution and transport it to the laboratory at 4°C;

[0029] (2) Use scissors to remove the subcutaneous fat tissue as much as possible;

[0030] (3) Disinfect the skin tissue treated in step (2) with iodophor for 10 minutes at room temperature;

[0031] (4) Wash the sterilized skin tissue 3 times with sterile PBS;

[0032] (5) Place the skin tissue cleaned in step (4) in a sterile petri dish, and cut into small pieces with a size of 5mm×5mm;

[0033] (6) Put the skin tissue from step (5) in 15ml of discrete enzyme, and store it overnight at 4°C.

[0034] 2. Preparation of amniotic membrane decellularized scaffold:

[0035] (1) Disinfection: After separating the amniotic membrane, put it into a centrifuge tube, add 20ml of alcohol with a volume fraction of 65-75% per 15g of amniotic me...

Embodiment 2

[0052] Embodiment 2 The present invention is realized through the following steps.

[0053] 1. Skin tissue preparation:

[0054] (1) Quickly place the excised autologous skin in sterile PBS solution and transport it to the laboratory at 4°C;

[0055] (2) Use scissors to remove the subcutaneous fat tissue as much as possible;

[0056] (3) Disinfect the skin tissue treated in step (2) with iodophor for 10 minutes at room temperature;

[0057] (4) Wash the sterilized skin tissue 3 times with sterile PBS;

[0058] (5) Place the skin tissue cleaned in step (4) in a sterile petri dish, and cut into small pieces with a size of 5mm×5mm;

[0059] (6) Put the skin tissue from step (5) in 15ml of discrete enzyme, and store it overnight at 4°C.

[0060] 2. Preparation of amniotic membrane decellularized scaffold:

[0061] (1) Disinfection: After separating the amniotic membrane, put it into a centrifuge tube, add 20ml of alcohol with a volume fraction of 65-75% per 15g of amniotic me...

Embodiment 3

[0078] Embodiment 3 The present invention is realized through the following steps.

[0079] 1. Skin tissue preparation:

[0080] (1) Quickly place the excised autologous skin in sterile PBS solution and transport it to the laboratory at 4°C;

[0081] (2) Use scissors to remove the subcutaneous fat tissue as much as possible;

[0082] (3) Disinfect the skin tissue treated in step (2) with iodophor for 10 minutes at room temperature;

[0083] (4) Wash the sterilized skin tissue 3 times with sterile PBS;

[0084] (5) Place the skin tissue cleaned in step (4) in a sterile petri dish, and cut into small pieces with a size of 5mm×5mm;

[0085] (6) Put the skin tissue from step (5) in 15ml of discrete enzyme, and store it overnight at 4°C.

[0086] 2. Preparation of amniotic membrane decellularized scaffold:

[0087] (1) Disinfection: After separating the amniotic membrane, add it to a centrifuge tube, add 20ml of alcohol with a volume fraction of 65-75% for each 15g of amniotic...

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Abstract

The invention discloses a preparation method of novel tissue engineering skin. The method includes steps of jointly culturing an amniotic membrane decellularized scaffold and a mesenchymal stem cell to be a carrier, inoculating horn cell; culturing by a serum-free culturing system, and acquiring a complete tissue engineering skin membrane. Through the method, the method can culture a complete skin membrane and the skin membrane can be applied to the skin scar repair and leucoderma treatment. The horn cell is originated from body skin or prepuce of the newborn. The carrier is low in immunogenicity while also has biological activity; the serum-free culturing system can realize the in-vitro serum-free cultivation and amplification of the horn cell, and improve cell growth, multiplication, cell form, activity and function instead of bad influence; the skin membrane cultivated by the method is approached to natural skin; after the skin-grafting culturing, it can be applied to the scar repair of scaled and other injured skins as well as treatment of skin disease such as leucoderma.

Description

technical field [0001] The invention relates to the field of tissue engineering skin construction and regenerative medicine, in particular to a novel tissue engineering skin preparation method for treating skin defects and repairing scars. Background technique [0002] Tissue-engineered skin is the most mature field of tissue engineering research. Its core content is to construct a three-dimensional scaffold that supports cell growth, and to co-culture with keratinocytes and / or fibroblasts in vitro to form a tissue that can be used for wound coverage and repair. skin equivalent. Among them, scaffold materials provide a space environment for seed cells to adhere, migrate, proliferate and differentiate, and play an important role in the construction of tissue engineered skin. [0003] Tissue engineered skin scaffold materials mainly include artificially synthesized tissue engineered skin scaffold materials (referred to as synthetic scaffold materials) and natural tissue engin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61L27/60A61L27/36A61L27/38
CPCA61L27/3633A61L27/3804A61L27/3834A61L27/3886A61L27/3895A61L27/60C12N5/0629C12N2500/90C12N2501/11C12N2501/30C12N2501/33C12N2533/90C12N2533/92
Inventor 张君祥姜大奎
Owner 林翠霞
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