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Method for preparing human body tissue engineering skin

A technology of engineering skin and human tissue, which is applied in the field of preparation of human tissue engineering skin, can solve the problems of high cost, complex matrix components, easy to cause immune diseases, etc., and achieve an easy-to-accept effect

Inactive Publication Date: 2005-04-20
刘凯
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But it has the following disadvantages: the ingredients used in the formation of the dermal matrix are diverse, including: pepsin, chitin, elastin, hyaluronic acid, chondroitin sulfate, heparan sulfate, and fibronectin, etc. There are many types of solutions, and the process It is very complicated, especially the use of human collagen, it is difficult to obtain materials, it is easy to cause immune diseases, the cost is expensive, it is not conducive to mass production and popularization, and there has been no clinical application report so far
Its matrix materials are collagen, chitin, chitin, gelatin, hyaluronic acid, chondroitin sulfate and polyglycolic acid. The cells are derived from mammalian skin. Although the cultured and freeze-dried skin can be preserved for a long time, it exists The disadvantages are as follows: the use of ammonia to neutralize the collagen gel produced is not conducive to the growth and development of cells; the ability to resist collagenase digestion and infection is poor; because the cell culture time is too short, the cultured artificial skin is not good in its structure. There is a large gap with natural skin, and the clinical application effect is not good; and the matrix composition is complex

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027]Embodiment 1: (A) 0.8 square centimeters of fresh isolated human skin (preserved in 0.1 molar phosphate buffered solution (PBS), pH7.4, 4 degrees, less than 24 hours) are transferred from phosphate buffered solution tube to containing 10 units of penicillin and streptomycin / ml in phosphate buffer tubes (20 ml). Shake for 2 minutes, repeat 2 times, and then change to a new tube containing only 20 ml of phosphate buffer. Then the tissue was taken out and placed in a petri dish, excess subcutaneous tissue was removed, transferred to a new petri dish, epidermal cell culture fluid (FAD-I) was added, placed in a 37 degree incubator, and cultivated for 20 hours. Afterwards, transfer them to a new petri dish, cut them into 0.05 mm2, evenly distribute them in the petri dish, and place them in a ventilated box for 1 hour to allow the tissues to dry and adhere to the bottom of the petri dish. Gently add epidermal cell culture solution (FAD-I), put it in a 37-degree incubator for c...

Embodiment 2

[0030] Example 2: (A) 1 square centimeter of freshly isolated human skin was transferred from a phosphate buffer tube to a phosphate buffer tube containing 10 units of penicillin and streptomycin / ml. Shake for 3 minutes, repeat 3 times, and then change to a new tube containing only 20 ml of phosphate buffer. Then the tissue was taken out and placed in a culture dish, excess subcutaneous tissue was removed, transferred to a new culture dish, epidermal cell culture fluid (FAD-I) was added, placed in a 37-degree incubator, and cultivated for 22 hours. Afterwards, transfer them to a new petri dish, cut them into pieces to a size of 0.08 mm2, distribute them evenly in the petri dish, and place them in a ventilated box for 1 hour to allow the tissues to dry and adhere to the bottom of the petri dish. Gently add epidermal cell culture medium, put it in a 37-degree incubator for culture, and change the culture medium once every 2 days. After about 2 days, the keratinocytes grow out, a...

Embodiment 3

[0033] Example 3: (A) 1.2 square centimeters of fresh isolated human skin was transferred from a phosphate buffer tube to a phosphate buffer tube containing 10 units of penicillin and streptomycin / ml. Shake for 4 minutes, repeat 4 times, and then change to a new tube containing only 20 ml of phosphate buffer. Then the tissue was taken out and put into a petri dish, and epidermal cell culture solution (FAD-I) was added, placed in a 37 degree incubator, and cultivated for 24 hours. Afterwards, transfer them to a new petri dish, cut them into pieces of 0.1 mm2, and evenly distribute them in the petri dish. After the tissues are dried, they adhere to the bottom of the petri dish. Add epidermal cell culture solution (FAD-I), put it in a 37-degree incubator for culture, and change the culture solution once every 2 days. After 3 days, keratinocytes grow out, and keratinocytes can be collected after 15 days. (B) Transfer 1.2 cm2 of freshly isolated human skin from a phosphate buffer ...

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Abstract

A method for preparing human tissue engineering skin comprises 1, preparation of cutin cell of human skin,2, culturing an preparation of fibroblastic cell, 3, preparation of epidermic cell culture solution and fibroblastic cell culture solution, 4, formation of matrix gel, 5, culturing compound skin, 6, preservation and conveying.

Description

Technical field [0001] The invention relates to a preparation method of human tissue engineered skin, which is cultivated from biologically active human epidermal keratin cells and dermal fibroblasts, and has a tissue structure very similar to human skin. It can be used clinically to treat refractory diabetic plantar ulcers, venous ulcers, burns, scalds and other skin wounds. Background technique [0002] According to the latest epidemiological survey data of the Diabetes Branch of the Chinese Medical Association, there are nearly 40 million diabetic patients in my country, and one of the common chronic complications is diabetic plantar ulceration, which can be healed with conventional debridement and saline dressing. Slow, low healing rate. And apply debridement and the artificial skin transplantation treatment of this invention, the wound healing time obviously shortens, and the healing rate obviously improves. In addition, the number of burn patients in our country is al...

Claims

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Application Information

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IPC IPC(8): A61L27/60C12N15/08
Inventor 刘凯
Owner 刘凯
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