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Method for fabricating activated artificial skin tissue in bilayer by using bioreactor

An artificial skin and active technology, applied in biochemical equipment and methods, microorganisms, tissue culture, etc., to achieve repeatable production, promote cell proliferation and functional expression, and reduce impact

Inactive Publication Date: 2005-01-12
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is to disclose a method for preparing double-layered active artificial skin tissue using a bioreactor, so as to overcome the above-mentioned deficiencies in the prior art and meet the needs of related fields

Method used

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  • Method for fabricating activated artificial skin tissue in bilayer by using bioreactor
  • Method for fabricating activated artificial skin tissue in bilayer by using bioreactor

Examples

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Embodiment 1

[0067] Take the foreskin of the child and cut it into 1cm 2 The left and right skin blocks were washed 3 times with PBS containing 200 U / mL gentamycin, 2 times with 75% alcohol, and 2 times with PBS. The skin tissue was added to 200 U / mL neutral protease (Sigma) and digested at 4°C for 18 hours to separate the dermis and epidermis. Keratinocytes are obtained from epidermal tissue and fibroblasts are obtained from dermal tissue.

[0068] Cut the epidermis into small pieces, act with 6 mL of 0.25% trypsin (Sigma) / 0.06% EDTA (the volume of trypsin / EDTA digestion solution is about 3 times the volume of the digested tissue) for 8 minutes, and add an equal amount of calf serum and trypsin activity.

[0069] Filter through a 150-mesh sieve to remove epidermal fragments and obtain keratinocyte suspension. Centrifuge at 2000r / min for 5min, discard the supernatant, wash twice with PBS, add keratinocyte serum-free medium (Gibco) containing 200U / mL gentamycin, and the cell inoculation ...

Embodiment 2

[0072] The fascia and lipids were removed from the tendon to make tendon powder, which was stored at -20°C for later use. Soak the tendon end in 0.1mol / L HAc solution to prepare a collagen-HAc solution with a final concentration of 8.0mg / mL. Chitosan solution, heparin sodium solution, and chondroitin sulfate solution were added to the collagen solution so that the final concentrations were 10% (g / g), 7% (g / g) and 3% (g / g) of the dry weight of collagen, respectively. g), frozen at -20°C overnight, and vacuum freeze-dried for 24 hours to obtain a collagen sponge. Collagen sponges were chemically cross-linked with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (Sigma), rinsed thoroughly with distilled water, freeze-dried, and sterilized with ethylene oxide to make usable Composite collagen sea surface matrix material for preparing double-layer active skin tissue.

Embodiment 3

[0074] The same method as in Example 2 is adopted, but in the composite collagen sponge, the amino polysaccharide substance is hyaluronic acid, and the final concentration is 20% (g / g) of the dry weight of the collagen.

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Abstract

A process for preparing artificial active dual-layer skin tissue by use of bioreactor includes putting the composite collagen sponge as scaffold on the porous supporter in bioreactor, adding fibroblast suspension to the bioreactor for configuring the dermis, inoculating horny cells, and culturing for configuring epiderm.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing an active artificial skin tissue with a double-layer structure of dermis and epidermis, in particular to a method for preparing an active artificial skin tissue by using a bioreactor. Background technique [0002] The skin is the largest organ of the human body, which plays a role in protecting the internal environment of the body and maintaining its stability. Because it is located on the body surface of the human body, it is susceptible to various injuries and diseases. Skin transplantation has been used to treat various acute and chronic skin diseases for thousands of years. The traditional treatment method is autologous or allogeneic transplantation. However, due to the limited source of autologous skin, allogeneic skin grafts have problems such as rejection, disease transmission, and difficulty in preservation. , it is difficult to meet the tra...

Claims

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Application Information

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IPC IPC(8): A61L27/60C12N5/08
Inventor 谭文松周燕
Owner EAST CHINA UNIV OF SCI & TECH
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