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Method for building tissue engineered skin by composite DED (de-epidermidalized dermis) of amnion endothelial cells

A technology for tissue engineering skin and amniotic epithelial cells, applied in medical science, prostheses, etc., can solve the problems of loss of proliferation ability, lack of basement membrane structure and function, etc., and achieve high application value

Inactive Publication Date: 2017-08-08
GUIZHOU PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although most of the current dermal substitutes can simulate the structure and function of the dermis, almost all lack effective basement membrane structure and function, and epidermal keratinocytes will gradually lose their ability to proliferate on this dermal substitute lacking basement membrane

Method used

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  • Method for building tissue engineered skin by composite DED (de-epidermidalized dermis) of amnion endothelial cells
  • Method for building tissue engineered skin by composite DED (de-epidermidalized dermis) of amnion endothelial cells
  • Method for building tissue engineered skin by composite DED (de-epidermidalized dermis) of amnion endothelial cells

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Embodiment 1

[0032] A method for constructing tissue-engineered skin with amnion epithelial cells combined with de-epidermized dermis, using human amnion epithelial cells (hAECs) as seed cells, using human de-epidermized dermis (De-epidermized dermis, DED) as a scaffold material, and seeding Cells are planted on the scaffold material to construct hAECs-DED tissue engineered skin, including the following steps: 1) isolation and culture of hAECs; 2) isolation and culture of fibroblasts (FB); 3) preparation of DED; 4) hAECs-DED tissue engineering The construction of the skin.

[0033] Wherein, the specific method of isolating and culturing the hAECs is as follows: the amnion tissue is washed repeatedly with phosphate buffered saline (PBS) and cut into small pieces, and then added with 0.05% trypsin-0.02% ethylenediaminetetraacetic acid (EDTA) Solution, digested at 37°C for 10min, discarded the digestate (mainly to remove miscellaneous cells and cell debris); then repeated the above method to ...

Embodiment 2

[0043] A method for constructing tissue-engineered skin with amnion epithelial cells combined with de-epidermized dermis, using human amnion epithelial cells (hAECs) as seed cells, using human de-epidermized dermis (De-epidermized dermis, DED) as a scaffold material, and seeding Cells are planted on the scaffold material to construct hAECs-DED tissue engineered skin, including the following steps: 1) isolation and culture of hAECs; 2) isolation and culture of fibroblasts (FB); 3) preparation of DED; 4) hAECs-DED tissue engineering The construction of the skin.

[0044] Wherein, the specific method of isolating and culturing the hAECs is as follows: the amnion tissue is washed repeatedly with phosphate buffered saline (PBS) and cut into small pieces, and then added with 0.05% trypsin-0.02% ethylenediaminetetraacetic acid (EDTA) Solution, digested at 37°C for 10min, discarded the digestate (mainly to remove miscellaneous cells and cell debris); then repeated the above method to ...

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Abstract

The invention discloses a method for building tissue engineered skin by composite DED (de-epidermidalized dermis) of amnion endothelial cells. The hAECs (human amnion endothelial cells) are used as seed cells; human DED is used as a stent material; the seed cells are planted onto the stent material to build the hAECs DED; the seed cells are planted onto the stent materials to build the hAECs-DED tissue engineered skin. The method comprises the following steps of (1) hAECs separation culture; (2) FB (fibroblast) separation culture; (3) DED preparation; (4) hAECs-DED tissue engineered skin building. The amnion endothelial cells are used as the seed cells; the human DED is used as a stent; the seed cells and the stent are all from the human body and have the homeology; the built tissue engineered skin has the tissue structures and functions similar to the normal skin, and also has some features of normal skin proliferation and differentiation.

Description

technical field [0001] The invention relates to the technical field of tissue-engineered skin, in particular to a method for constructing tissue-engineered skin by combining amniotic epithelial cells with de-epidermal dermis. Background technique [0002] In recent years, various tissue-engineered skins such as Apligraf VR and Dermagraft VR have been applied clinically to repair wounds. Most of the seed cells of tissue-engineered skin are allogeneic keratinocytes, but considering many aspects, keratinocytes are not ideal seed cells. In order to make tissue-engineered skin better clinically, using amniotic epithelial cells (hAECs) as seed cells will be a better and more innovative choice. The advantage of hAECs over other cells is that it has the plasticity to differentiate into three germ layers, and it expresses growth factors (EGF, KGF, HGF) and anti-inflammatory and antibacterial cytokines, which are beneficial to wound healing. Because amniotic membrane is postpartum w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/60A61L27/38A61L27/36
CPCA61L27/362A61L27/3687A61L27/3804A61L27/3813A61L27/60
Inventor 江蕾微陆洪光聂瑛洁盛良马婷时光
Owner GUIZHOU PROVINCIAL PEOPLES HOSPITAL
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