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Production method for cryopreserved acellular dermal matrix, and cryopreserved acellular dermal matrix produced thereby

A technology of cryopreservation and dermal matrix, applied in biochemical equipment and methods, preservation of human or animal bodies, skin diseases, etc., can solve problems such as fast decomposition, destruction of collagen tissue, poor supply and demand, etc., and achieve tissue stability sex high effect

Active Publication Date: 2012-07-11
IND ACADEMIC COOP FOUND HALLYM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the price of this kind of skin for transplantation is very expensive, and most of them are imported products, so there is a problem of poor supply and demand
Using the skin of a donor’s cadaver, after removing the epidermis, in order to facilitate preservation, the acellular dermal matrix from which the inner cells of the dermis have been removed is often frozen and then used to eliminate immune rejection. However, during the freezing process, the acellular dermal matrix The collagen tissue inside is destroyed, and there is a problem of rapid decomposition after transplantation

Method used

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  • Production method for cryopreserved acellular dermal matrix, and cryopreserved acellular dermal matrix produced thereby
  • Production method for cryopreserved acellular dermal matrix, and cryopreserved acellular dermal matrix produced thereby
  • Production method for cryopreserved acellular dermal matrix, and cryopreserved acellular dermal matrix produced thereby

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0030] Prepare skin for cryopreservation using porcine skin as described below.

[0031] (1) Wash the pigskin with normal saline.

[0032] (2) Cut the pork skin into 5×10cm 2 the size of.

[0033] (3) Pig skin was added to 1M NaCl (Sigma, USA) solution.

[0034] (4) Prepare a reactor (P-039, KoaTech) at 38°C.

[0035] (5) The pig skin placed in a 1M NaCl (Sigma, USA) solution was subjected to a stirring reaction in a reactor at 38° C. for about 6 to 24 hours.

[0036] (6) Use medical tweezers to remove the epidermis.

[0037] (7) The dermis from which the epidermis was removed was washed with a phosphate buffer solution (pH 7.2, GIBCO, USA).

[0038] (8) Put the washed dermis into 0.1% SDS, stir and react at room temperature for 1 hour, so as to remove the inner cells of the dermis.

[0039] (9) The dermis from which cells were removed was washed with a phosphate buffer solution.

[0040] (10) Glycerol (Sigma, USA) and phosphate buffer were mixed at a weight ratio of 1:...

experiment example 1

[0067] Experimental Example 1 Histological identification

[0068] H&E staining was carried out on the pigskins prepared in the above-mentioned examples and comparative examples according to the following method.

[0069] (1) A paraffin block was cut into a thickness of 4 μm, and then dried to make a paraffin section.

[0070] (2) implement dewaxing process, react 3 times in xylene, each 5 minutes; React 3 times in 100% ethanol, each 2 minutes; React 1 time, 1 minute in 90% ethanol; React once in % ethanol for 1 minute; react once in 70% ethanol for 1 minute; then wash with running water for 10 minutes.

[0071] React in Hematoxylin staining solution for 10 minutes, then wash with flowing water for 3 minutes, react in Eosin (Eosin) staining solution for 10 minutes, and then wash with flowing water until no eosin staining solution flows out . React 10 times in 70% ethanol, 1 second each; react 10 times in 80% ethanol, 1 second each; react 10 times in 90% ethanol, 1 second ...

experiment example 2

[0081] Experimental Example 2 Measurement of decomposing properties by collagen hydrolase

[0082] In order to observe the change of the stability of the acellular dermal matrix with the difference of the sucrose concentration, the decomposing property of the collagen decomposing enzyme was measured according to the following procedure.

[0083] (1) 25 mg of the sample was added to 5 mM TES buffer mixed with 5 ml of 0.36 mM calcium chloride (calcium chloride) and mixed uniformly.

[0084] (2) Add 0.1 ml of 0.1 mg / ml collagenase (collagenase) to the sample of (1), mix well, and react at 37° C. for 1 day.

[0085](3) After serial dilution with 4.0mM L-leucine standard solution (L-leucine standard solution), treat with ninhydrin col or reagent, and measure at 570nm Absorbance (VERSA max, Molecular Device, USA) was determined and a standard curve was drawn.

[0086] (4) The sample of (2) was treated with a ninhydrin developer, and the absorbance was measured at 570 nm.

[0087]...

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Abstract

The present invention relates to a production method for cryopreserved acellular dermal matrix and to cryopreserved acellular dermal matrix produced thereby, and more specifically it relates to a method in which a cryopreservation agent is made by adding sucrose to basic components consisting of glycerol and a basic solution and in which the resulting solution is used in the cryopreservation of skin tissue from which the cells in the epidermis and dermis have been removed, and relates to cryopreserved acellular dermal matrix produced thereby.

Description

technical field [0001] The invention relates to a preparation method of cryopreserved acellular dermal matrix (ADM) and a cryopreserved acellular dermal matrix prepared therefrom. In more detail, it relates to a method of using glycerin and a basic solution as basic components, adding sucrose to prepare a cryoprotectant, and then using the solution to cryopreserve skin tissue from which epidermis and dermis cells have been removed, and its Preparation of acellular dermal matrix for cryopreservation. Background technique [0002] The skin is the largest organ covering the entire body surface. It prevents the loss of body fluids, prevents the invasion of harmful substances and microorganisms, resists physical and chemical stimuli, etc., and performs the function of protecting our body. For patients with severely damaged skin due to severe burns, trauma, epithelial cancer resection, and skin diseases, it should act as a protective film to prevent infection of the damaged area ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/60A61L27/36A61L27/38C12N5/02
CPCA61L2430/40A61L27/362A61L27/60A01N1/0221A61L27/3687A61P17/00A61L27/36A61L27/38C12N5/00
Inventor 全旭崔源益朴晚成金根亨郑载得
Owner IND ACADEMIC COOP FOUND HALLYM UNIV
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