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Novel method for constructing tissue engineering skin

A tissue-engineered skin and new method technology, applied in medical science, prosthesis, etc., can solve problems such as affecting the adhesion and proliferation of epidermal stem cells, delaying the healing speed of defect wounds, lack of fibroblasts, etc. The effect of low immunogenicity, good biocompatibility

Active Publication Date: 2014-06-18
CHINA INST FOR RADIATION PROTECTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are some deficiencies in this method. One is that the scaffold material used only undergoes epidermis removal without decellularization, which will cause the scaffold material to contain more cellular components, which will easily lead to the occurrence of immune rejection when used in clinical practice; Using epidermal stem cells as seed cells, lack of fibroblasts, affects the adhesion and proliferation of epidermal stem cells, and will also delay the healing speed of defect wounds

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] (1) Isolation and culture of epidermal stem cells

[0033] Take 2~4cm 2 Same kind of skin tissue (get 3cm in the present embodiment 2 ), placed in double-antibody-containing PBS for repeated washing, cut off fat and subcutaneous tissue, and cut into 0.5×0.5cm 2 The small pieces were transferred into Dispase II (2.4U / ml) and kept overnight at 4°C. Separate the dermis and epidermis, collect the epidermis slices and cut them into pieces, digest them in 0.25% trypsin and 0.02% EDTA mixed digestion solution at 37°C for 30 minutes, terminate the serum, and filter through a 200-mesh cell sieve. Centrifuge at 1000rpm for 5 minutes, discard the supernatant, resuspend in the special medium for epidermal stem cells, and adjust the cell density to (1-2)×10 6 ml, inoculated in type IV collagen-coated culture flasks, and after rapid screening for 15 minutes, the non-adherent cells were discarded and replaced with new culture based on CO 2Cultivate in an incubator, change the medi...

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Abstract

The invention provides a method for constructing tissue engineering skin. In the method, human epidermal stem cells and hypodermal fibroblasts are taken as seed cells, and a homologous acellular dermal matrix modified by a human placental type IV collagen is taken as a stent, so that active dual-layer engineering skin is compounded and constructed by using a gas-liquid interface separate culture method. The tissue engineering skin constructed by using the method is closer to the structure of natural skin; and as proved by histomorphology, the tissue engineering skin has an epidermal and hypodermal double-layer structure, wherein the hypodermal collagen stent has a complete structure, an epidermal layer is provided with a plurality of layers of cells of different differentiation degrees, and the morphological requirement of the tissue engineering skin is met.

Description

technical field [0001] The invention belongs to human tissue engineering technology, and specifically relates to a new method for constructing tissue-engineered skin. Human epidermal stem cells and dermal fibroblasts are used as seed cells, and the same acellular dermal matrix modified by human placenta type IV collagen is used as a scaffold. A kind of active bilayer tissue engineered skin was compositely constructed by air-liquid interface separation method. Background technique [0002] The skin is the largest tissue and organ of the human body and the outer barrier of the human body. The integrity of the skin plays an important role in human beings. Burns (trauma) injuries and other causes of large-area skin defects can lead to serious dysfunction of the human body, and even life-threatening. At present, the most effective method for repairing skin defects is autologous skin transplantation, but autologous skin sources for large-area burns often cannot meet the needs of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/60A61L27/40
Inventor 李宝兴董丽李宝明马绍英李靖张乃丽周沫王旭昇刘晓明张育敏陈学英李幼忱赵亚平
Owner CHINA INST FOR RADIATION PROTECTION
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