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Culture method for acquisition of tissue-engineered epidermis and application of culture method

A technology of tissue engineering and culture method, which is applied in artificial cell constructs, medical science, prostheses, etc. It can solve the problems that all kinds of cells cannot be guaranteed to grow well, the structure of natural skin is different, and the metabolic activities are not clear.

Active Publication Date: 2015-12-09
NOVAPRINT THERAPEUTICS SUZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This tissue-engineered composite skin is derived from autologous cells, has no rejection reaction, and has certain strength and toughness due to the existence of the scaffold, but there are still some defects: 1. There is a competitive relationship between keratinocytes and fibroblasts when they are co-cultured, Therefore, the growth state of several kinds of cells when they grow together on the biological scaffold may exist that one kind of cell inhibits another kind of cell, which cannot guarantee the good growth of all kinds of cells; The growing full-thickness skin has some contact problems such as adhesion; 3. Whether the environmental changes after transplantation can make the cells on the scaffold survive well and various metabolic activities are still unclear
[0004] For other allogeneic skin tissue engineering products, there are still differences in the structure of natural skin. The rejection of allogeneic skin cells, the survival of cells on the biological scaffold, the growth state and the secretion of the cell matrix will be different. Problems such as rapid integration with the transplanted surface

Method used

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  • Culture method for acquisition of tissue-engineered epidermis and application of culture method
  • Culture method for acquisition of tissue-engineered epidermis and application of culture method
  • Culture method for acquisition of tissue-engineered epidermis and application of culture method

Examples

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example 1

[0064] Example 1: Successful cultivation of epidermal patches using adult breast skin.

[0065] (1) Materials and methods

[0066] Sample: About 3cm of adult normal chest skin discarded by hospital surgery 2

[0067] Skin medium: KGM-Gold medium, DMEM / F12 mixed medium

[0068] (2) Cell isolation and skin slice culture

[0069] a) The skin sample is refrigerated and transported to the laboratory in DMEM culture medium, and the area is measured;

[0070] b) Clean the sample with PBS; repeat the cleaning of the sample after disinfection with iodine tincture and alcohol, cut off 2 / 3 of the dermis and chop it;

[0071] c) 0.05% trypsin (DPBS and 0.25% trypsin / EDTA according to the ratio of 4:1) digested the sample cold for 12-16 hours; add trypsin inhibitor to stop the digestion;

[0072] d) Type I collagenase at a concentration of 0.05%-0.5% continues to be thermally digested for 2-6 hours, after filtering, centrifuging, collecting epidermal cells and inoculating them in cult...

example 2

[0080] Example 2: Application of autologous epidermal membrane transplantation in repairing burn scars.

[0081] Skin slice culture and quality inspection: the specific steps refer to the description in Example 1;

[0082] Skin grafting: on the patient's scar site (such as Figure 3-1 , hypopigmented leukoplakia) was disinfected with medical alcohol, and the epidermis and scar tissue were removed with a surgical water knife. Spread the epidermal membrane sheet obtained in Example 1 on the surgical site. Cover with sterile gauze and secure with a bandage.

[0083] Results: After 3 months of transplantation, the first follow-up was conducted to observe the scar repair at the transplanted site. The tissue-engineered epidermis was well integrated with its own tissue. Because some parts were cut too deeply during the operation, hypertrophic scars were formed, which were slightly darker than other parts (such as Figure 3-2 ), but as time goes on, it gradually returns to the same...

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Abstract

The invention provides a culture method for acquisition of tissue-engineered epidermis and application of the culture method. By means of co-culture of epidermal cells and melanocytes, an epidermis membrane with a stratum bassale, a stratum spinosum, a stratum granulosum and a stratum corneum can be acquired, and large area, avoidance of rejection reaction, structural similarity to natural epidermis, excellent treatment effects and the like are realized.

Description

technical field [0001] The invention relates to a method for culturing and obtaining tissue-engineered epidermis, in particular to a method for culturing the epidermis by co-cultivating melanocytes and keratinocytes. Background technique [0002] Tissue-engineered skin has more and more applications in the medical and cosmetic fields as the technology matures, and is far superior to other treatments in terms of safety and extensive treatment effects. In the treatment of skin tissue diseases, besides drug therapy, two treatment methods are mostly implemented: direct autologous skin transplantation and allogeneic skin or skin substitute transplantation; autologous grafts cannot complete the treatment of a large area at one time, requiring repeated operations . [0003] In products that can treat a large area, autologous or allogeneic epidermal cells are often extracted and cultured to obtain products. Now there is a patent for inoculating the extracted epidermal cells and fib...

Claims

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Application Information

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IPC IPC(8): C12N5/071A61L27/38A61L27/60
Inventor 刘洪李琳杨熙
Owner NOVAPRINT THERAPEUTICS SUZHOU CO LTD
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