Novel human epidermal culture method

A culture method and epidermis technology, applied in the direction of artificial cell constructs, vertebrate cells, animal cells, etc., can solve problems such as unfavorable quality control, increased risk of infection and immune response, unfavorable clinical use of cultured epidermis, etc., to reduce costs and unsafe factors, reducing the effect of mixing

Inactive Publication Date: 2016-01-06
赫柏慧康生物科技无锡有限公司
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  • Summary
  • Abstract
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  • Application Information

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Problems solved by technology

Because the use of serum and trophoblast cells from animals is not conducive to the clinical use of cultured epidermis, increases the risk of infection and immune response, and is not conducive to quality control

Method used

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Embodiment Construction

[0038] Preferred embodiments of the present invention are described below, and it should be understood that the preferred embodiments described here are only used to illustrate and explain the present invention, and are not intended to limit the present invention.

[0039] A novel method for culturing human epidermis, comprising the following steps:

[0040] (1) Skin sample processing and cell separation:

[0041] Take 3cm of the patient's skin 2 Store in DMEM (Gibco); disinfect the patient's skin with ethanol to remove microbial contamination on the patient's skin surface; rinse the patient's skin several times with phosphate buffer solution to remove residual ethanol on the patient's skin, and cut the patient's skin with a scalpel into tiny particles with a diameter of 2mm;

[0042] Put the fine particles of the cut patient's skin into a PE bottle filled with 40ml of collagenase at a concentration of 100-200U / ml, and use a cell filter to collect the epidermal tissue after ...

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Abstract

The invention discloses a novel human epidermal culture method. The method includes the following steps of (1), skin sample treatment and cell separation; (2), culture of epidermal cells; (3), passage of the epidermal cells and stripping of tissue engineered skin; (4), quality inspection of the tissue engineered skin. A preparation method of a KMSFM culture medium includes: taking DSFM of Gibco as a basis; using low calcium ion concentration; adding bovine pituitary extract (BPE), bovine insulin, bovine transferrin, bFGF, cortisol, heparin and endothelin at the same time. By the novel human epidermal culture method, bovine serum is removed, mixing of non-human-derived ingredients can be reduced greatly, and cost and unsafe factors are reduced; a culture system coated by I-type collagen is used, cell growth slowness caused by removing of nourishing layer cells is avoided, and the novel human epidermal culture method can be used for treating skin pigment diseases such as leukoderma and giant pigmented nevus.

Description

technical field [0001] The invention relates to a novel method for culturing human epidermis and tissue engineered skin prepared therefrom. Background technique [0002] The skin is the largest organ of the human body, and it consists of three parts, namely the epidermis, dermis and subcutaneous layer. The epidermis is located in the outermost layer of the skin, with a thickness between 0.06-0.8mm, and its microstructure is composed of basal layer, spinous cell layer, granular cell layer and stratum corneum. The "Green method" of epidermal culture technology is a multilayer culture method of skin epidermal cells established by Professor Howard Green of Harvard Medical School in 1975. [0003] The "Green method" method refers to the cultivation of several layers of cells, which eventually form an epidermis. This method successfully produced epidermal tissue similar to skin. This structure similar to the normal epidermis is an aggregate of keratinocytes, which is formed by ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 刘洪李琳杨熙
Owner 赫柏慧康生物科技无锡有限公司
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