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Tissue engineering epidermis substitute having pigment and its preparation method

A technology of tissue engineering and substitutes, which is applied in the field of preparation of pigmented artificial epidermal substitutes, which can solve the problems of inability to repair pigment cells, cell membrane transplantation and inaccurate curative effect of melanocyte injection, etc., and achieve the perfect effect of skin repair

Inactive Publication Date: 2006-06-14
吴莲莲 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this kind of epidermal substitute constructed solely with epidermal cells cannot repair the lost pigment cells in the damaged site during epidermalization. Loss of pigment left at the damaged site in the future
In addition, there are a large number of depigmentation patients (such as vitiligo) in clinical practice. The conventional treatment methods for these patients include skin grafting, cell sheet transplantation, and injection of melanocytes. However, skin grafting brings new wounds to patients, and cell sheet transplantation and The curative effect of melanocyte injection is uncertain, therefore, there is an urgent need for a new and effective treatment method in clinical practice

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0013] 1. Isolation and culture of epidermal cells and melanocytes:

[0014] Wash the skin tissue with chloramphenicol solution for 3-5 times, or (and) soak with 75% ethanol solution for 1 minute, and wash with PBS for 2-3 times; Digest with 0.5u / ml Dispase Digest at 37°C for 2-7 hours, or digest at 4°C for 8-12 hours, separate the epidermis and dermis; then add 0.05% trypsin and 0.53mM EDTA to digest the epidermis for 8-20 minutes , stop digestion with trypsin inhibitor, filter through 150-mesh stainless steel mesh, collect cells and divide them into two parts, one part is cultured with epidermal cell culture medium, the other part is cultured with melanocyte culture medium, and cell expansion is carried out separately .

[0015] Melanin medium is commercialized M154 medium plus BPE (0.2% v / v), insulin (5 μg / ml), transferrin (5 μg / ml), basic fibroblast growth factor (3ng / ml), hydrogenation Cortisone (0.18μg / ml), heparin (3μg / ml), TPA (10ng / ml).

[0016] 2. Preparation of b...

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Abstract

The invention relates to band pigment labor epidermis substitute manufacturing method and its application. Its feature is that the used cell contains epidermis cell and melanocyte. Its manufacturing method are dissolving chitosan by 1% acetic acid; dissolving gelatine by distilled water; mixing them; heating them on plane dish, drying to form film material; taking it out; cleaning and dipping by a lot of distilled water; heating; setting in culture solution; setting the upper cell-film material compound in the culture solution. Compared with the current technique, the invention can repair epithelial tissue of the wound par, at the same time repair the melanocyte.

Description

[technical field] [0001] The invention relates to the field of tissue engineering, in particular to a preparation method and application of a pigmented artificial epidermis substitute. [Background technique] [0002] The skin is the largest organ of the human body and plays an important role in protecting the body. When skin defects are caused by various reasons, it will cause obstacles to the function of the body, and even life-threatening. For skin tissue defects, the common clinical treatment methods are split skin grafts or full-thickness skin grafts and vaseline gauze. Among them, split skin grafts are recognized standard treatment methods, but there are new problems caused by the existence of this treatment method. Skin trauma, and shortage of donors. [0003] After skin defect, the hypertrophic scar at the defect site is an important complication. Studies have shown that if the skin defect site can be quickly epithelialized, the scar formation at the defect site can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61F2/10A61L27/60C12N5/071
Inventor 吴莲莲丁建明
Owner 吴莲莲
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