Reagent for acellular processing of animal skin tissue, and processing method thereof

A technology for animal skin and treatment methods, applied in the direction of animal cells, vertebrate cells, artificial cell constructs, etc., to achieve the effects of suitable promotion, good biocompatibility, and low immune activity

Inactive Publication Date: 2013-12-11
CHONGQING ZONGSHEN JUNHUI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method also removes cells more thoroughly, and the basement membrane remains intact, but the content of type IV c

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Weigh 2.51g of SDS, 0.4g of NaOH and dissolve in 1L of distilled water, shake to fully dissolve and set aside.

[0033] A. SDS NaOH decellularization treatment

[0034] ① Press the animal skin to 10cm 2 Add to the container containing 50ml of SDS NaOH;

[0035] ②. Put the container in a constant temperature shaking incubator and shake it for 1 hour at 25°C;

[0036] ③. Wash with purified water for 1 hour, and change the liquid every 2 hours.

[0037] B. Glutaraldehyde crosslinking

[0038] ①. Add the skin sheet in step A to a 50ml container of glutaraldehyde cross-linking solution with a concentration of 0.01%;

[0039] ②. Put the container in a constant temperature shaking incubator and shake it for 1 minute at 25°C;

[0040] ③. Add glycine with a concentration of 0.01M / L to terminate the cross-linking reaction;

[0041] ④. Wash with purified water for 1 hour, and change the liquid every 2 hours.

[0042] C. Sterilize by irradiation with a dose of 1Kgy to obtain...

Embodiment 2

[0044] Weigh 28.838g of SDS, 4.0g of NaOH and dissolve in 1L of distilled water, shake to fully dissolve and set aside.

[0045] A. SDS NaOH decellularization treatment

[0046] ① Press the animal skin to 100cm 2 Add to the container containing 200ml of SDS NaOH;

[0047] ②. Put the container in a constant temperature shaking incubator and shake it at 37°C for 24 hours;

[0048] ③. Wash with purified water for 24 hours, and change the liquid every 2.5 hours.

[0049] B. Glutaraldehyde crosslinking

[0050] ①. Add the skin sheet in step A to 200ml of 0.1% glutaraldehyde cross-linking liquid container;

[0051] ②. Put the container in a constant temperature shaking incubator and shake it for 60 minutes at 37°C;

[0052] ③. Add glycine with a concentration of 0.1M / L to terminate the cross-linking reaction;

[0053] ④. Wash with purified water for 1 hour, and change the liquid every 2 hours.

[0054] C. Sterilize by irradiation with a dose of 20Kgy to obtain ADM.

Embodiment 3

[0056] Weigh 288.38g of SDS, 40g of NaOH and dissolve in 1L of distilled water, shake to fully dissolve and set aside.

[0057] A. SDS NaOH decellularization treatment

[0058] ① Press the skin to 200cm 2 Add to the container containing 300ml of SDS NaOH;

[0059] ②. Put the container in a constant temperature shaking incubator and shake it at 42°C for 48 hours;

[0060] ③. Wash with purified water for 48 hours, and change the liquid every 3 hours.

[0061] B. Glutaraldehyde crosslinking

[0062]①. Add the skin sheet in step A to a 300ml container of glutaraldehyde cross-linking solution with a concentration of 1.0%;

[0063] ②. Put the container in a constant temperature shaking incubator and shake it at 42°C for 120 minutes;

[0064] ③. Add glycine with a concentration of 1M / L to terminate the cross-linking reaction;

[0065] ④ Wash with purified water for 48 hours and change the solution every 3 hours.

[0066] C. Sterilize by irradiation with a dose of 35Kgy to obta...

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Abstract

The invention relates to the technical field of acellular processing of an animal skin tissue in biomedical engineering, and particularly relates to a reagent for acellular processing of a animal skin tissue, and a processing method thereof. The reagent is a solution prepared from SDS (sodium dodecyl sulfonate), NaOH and water. The processing method comprises the following steps of: A, acellular processing of SDS.NaOH; B, glutaraldehyde cross-linking; and C, sterilization. In the technical scheme disclosed by the invention, the animal skin tissue is directly processed by adopting the solution containing the SDS and the NaOH to obtain an acellular dermal matrix. Compared with other methods, the processing method provided by the invention has the characteristics of being simple and convenient, efficient, cheap, suitable for popularization and the like. The dermal matrix processed by the method is thorough in cell removal, a basement membrane complex is completely reserved, and the contents of collagen IV, fibronectin, elastin, HLA-DR (human leukocyte antigen DR) and the like in derma are very low.

Description

technical field [0001] The invention relates to the technical field of decellularization of animal skin tissue in biomedical engineering, in particular to a solution for decellularization of animal skin tissue and a treatment method thereof. Background technique [0002] In recent years, acellular dermal matrix (acellular dermalma-trix, ADM) as a new dermal substitute has gradually attracted people's attention. An ideal ADM should have: the cell components and type I and type II cytocompatibility antigens have been completely eliminated, the basement membrane complex (basement membrane complex. The generated specific cellular immune response (rejection reaction) will not induce non-specific foreign body reaction, and has good biocompatibility. Studies have shown that the immunogenicity of allogeneic dermis is mainly due to the immune response stimulated by the epidermal cells of the donor skin, fibroblasts in the dermis, endothelial cells and other cellular components, and ...

Claims

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Application Information

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IPC IPC(8): C12N5/071A61L27/36A61L27/60
Inventor 苟元彬张玉兰盖增周桂安牛春艳
Owner CHONGQING ZONGSHEN JUNHUI BIOTECH
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