Preparation of artificial skin containing hair follicle and artificial skin prepared by the same

Abstract

The invention relates to a preparation method of artificial skin, in particular to a preparation method of artificial skin with hair follicles and the skin made by the method, belonging to biomedical field. The preparation method comprises the following steps: (1) preparing the epithelial cells of the hair follicle; (2) culturing the dermal papilla cells; (3) culturing the artificial skin with hair follicles. The preparation method of artificial skin has the advantages that: (1) the invention originally prepares artificial skin with hair follicles on skin support by hair follicle cells, and reduces the antigenicity greatly compared with the artificial skin made by animal cells; (2) the artificial skin comprises hair follicles, and enable to do physiological functions of skin; (3) the artificial skin is not easy to shrink but easy to absorb; (4) the artificial skin can be used as wound-cover material for large area burn wounded person in clinic and accelerates the wound to heal naturally; (5) the invention resolves the trouble of skin source for large area burn; (6) the preparation method is reasonable in design, convenient in operation and easy in scale production.

Description

Technical field

[0001] The invention belongs to biomedicine, and relates to a preparation method of artificial skin, in particular to a preparation method of artificial skin containing hair follicles and the artificial skin prepared by the method. Background technique

[0002] Recently, artificial skins containing dermis and epidermis have been successfully developed at home and abroad, but these artificial skins have no other cellular components. For example, there are no sebaceous gland cells. The product secreted by sebaceous glands is called sebum, which has the effect of lubricating the skin and forms on the skin surface. Lipid membranes form part of the skin barrier. The free fatty acids in the lipid membrane inhibit the growth of certain pathogenic microorganisms; the fibroblasts in the dermal tissue around the hair follicle can produce natural antibiotics and have antibacterial defense capabilities. The outer hair root sheath in the hair follicle is equivalent to the spi...

Examples

Embodiment 1

[0031] (1) Preparation of hair follicle epithelial cells: Isolate human scalp hair follicles, add a certain concentration of 0.5% dispase enzyme (dispase, Sigma), digest for 12 to 18 hours at 4°C, transfer the hair follicles into D-Hanks buffered salt solution, and rinse Remove the separating enzyme, squeeze the hair shaft from the hair follicle, digest it with 0.15% trypsin plus 0.02% EDTA (Sigma) into individual hair follicle epithelial cells, centrifuge to remove trypsin, use serum-free keratinocyte culture medium, containing epidermal growth factor (EGF, Sigma-aldrich company) 10ng / mL, hydrocortisone 0.4μg / mL, insulin 5μg / mL, culture for use.

[0032] (2) Hair papilla cell culture: The human hair follicles from which the hair shaft has been removed are digested with 0.1-0.5% collagenase VI at 37°C for 2-6 hours. The dermal sheath surrounding the hair follicle is digested into single cells and the hair papilla inside it When the digestion has not yet started, stop the digestio...

Embodiment 2

[0036] The subcultured dermal papilla cells (4 / ml. Other technical solutions are as in Example 1. After 6 weeks of culture, H.E. staining was performed, and the formation of hair follicle-like structures was seen.

Embodiment 3

[0038] The subcultured dermal papilla cells (5 / ml. Other technical solutions are as in Example 1. After 6 weeks of culture, H.E. staining was performed, and the formation of hair follicle-like structures was seen.

Technical field

[0001] The invention belongs to biomedicine, and relates to a preparation method of artificial skin, in particular to a preparation method of artificial skin containing hair follicles and the artificial skin prepared by the method. Background technique

[0002] Recently, artificial skins containing dermis and epidermis have been successfully developed at home and abroad, but these artificial skins have no other cellular components. For example, there are no sebaceous gland cells. The product secreted by sebaceous glands is called sebum, which has the effect of lubricating the skin and forms on the skin surface. Lipid membranes form part of the skin barrier. The free fatty acids in the lipid membrane inhibit the growth of certain pathogenic microorganisms; the fibroblasts in the dermal tissue around the hair follicle can produce natural antibiotics and have antibacterial defense capabilities. The outer hair root sheath in the hair follicle is equivalent to the spi...

Examples

Embodiment 1

[0031] (1) Preparation of hair follicle epithelial cells: Isolate human scalp hair follicles, add a certain concentration of 0.5% dispase enzyme (dispase, Sigma), digest for 12 to 18 hours at 4°C, transfer the hair follicles into D-Hanks buffered salt solution, and rinse Remove the separating enzyme, squeeze the hair shaft from the hair follicle, digest it with 0.15% trypsin plus 0.02% EDTA (Sigma) into individual hair follicle epithelial cells, centrifuge to remove trypsin, use serum-free keratinocyte culture medium, containing epidermal growth factor (EGF, Sigma-aldrich company) 10ng / mL, hydrocortisone 0.4μg / mL, insulin 5μg / mL, culture for use.

[0032] (2) Hair papilla cell culture: The human hair follicles from which the hair shaft has been removed are digested with 0.1-0.5% collagenase VI at 37°C for 2-6 hours. The dermal sheath surrounding the hair follicle is digested into single cells and the hair papilla inside it When the digestion has not yet started, stop the digestio...

Embodiment 2

[0036] The subcultured dermal papilla cells (4 / ml. Other technical solutions are as in Example 1. After 6 weeks of culture, H.E. staining was performed, and the formation of hair follicle-like structures was seen.

Embodiment 3

[0038] The subcultured dermal papilla cells (5 / ml. Other technical solutions are as in Example 1. After 6 weeks of culture, H.E. staining was performed, and the formation of hair follicle-like structures was seen.

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