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 method for preparing artificial skin, in particular to a method for preparing 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 cell components, such as no sebaceous gland cells, and the product secreted by sebaceous glands is called sebum, which has the effect of lubricating the skin and forms on the skin surface Fatty membrane, which forms part of the skin barrier. The free fatty acids in the lipid film can inhibit the growth of some pathogenic microorganisms; the fibroblasts in the dermal tissue around the hair follicles can produce natural antibiotics and have antibacterial defense capabilities. The outer hair root sheath in the hair follicle is equivalent to the spinous laye...

Examples

Embodiment 1

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

[0032] (2) Dermal papilla cell culture: Human hair follicles with hair shafts removed are digested with 0.1-0.5% collagenase VI at 37°C for 2-6 hours, and the dermal sheath surrounding the hair follicles is digested into single cells and the dermal papilla in it When the digestion has not yet started, stop the digestion,...

Embodiment 2

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

Embodiment 3

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

technical field

[0001] The invention belongs to biomedicine and relates to a method for preparing artificial skin, in particular to a method for preparing 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 cell components, such as no sebaceous gland cells, and the product secreted by sebaceous glands is called sebum, which has the effect of lubricating the skin and forms on the skin surface Fatty membrane, which forms part of the skin barrier. The free fatty acids in the lipid film can inhibit the growth of some pathogenic microorganisms; the fibroblasts in the dermal tissue around the hair follicles can produce natural antibiotics and have antibacterial defense capabilities. The outer hair root sheath in the hair follicle is equivalent to the spinous laye...

Examples

Embodiment 1

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

[0032] (2) Dermal papilla cell culture: Human hair follicles with hair shafts removed are digested with 0.1-0.5% collagenase VI at 37°C for 2-6 hours, and the dermal sheath surrounding the hair follicles is digested into single cells and the dermal papilla in it When the digestion has not yet started, stop the digestion,...

Embodiment 2

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

Embodiment 3

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

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