Process for preparing artificial active skin of skin stem cell collagen sponge film

Abstract

The preparation process of artificial active skin includes planting separated and purified skin stem cell to both up and down sides of collagen sponge film, in vitro inducing the up skin stem cell to differentiate into the epiderm layers, and in vitro inducing the down skin stem cell to differentiate into the corium tissue, so as to form artificial active skin tissue with epiderm and corium tissue structure. The naked Balb / C mouse experiment shows that the artificial active skin tissue may be used in promoting features that epiderm growth and tissue repairing in the wound. The present invention has the advantages of simple process and low cost, and the artificial active skin of the present invention is one kind of excellent skin substitute.

Description

technical field

[0001] The invention relates to artificial skin, in particular to a method for preparing skin stem cell collagen sponge membrane artificial active skin. Background technique

[0002] Tissue-engineered artificial skin is the first relatively mature tissue engineering product to be developed for commercial application. Currently, its representative products are Biobrane TM , Integra TM 、Dermagraft TM and Apligraft TM They have played a certain role in the treatment of diseases such as burns, chronic venous ulcers and congenital skin deformities. However, to date, no product has been constructed using human skin stem cells as seed cells. Contents of the invention

[0003] The object of the present invention is to provide a method for preparing artificial active skin using human skin stem cells as seed cells for repairing, maintaining and improving skin tissue functions, simple preparation method and low cost skin stem cell collagen sponge membrane . [...

Examples

Embodiment Construction

[0024] The present invention is described in further detail below in conjunction with embodiment.

[0025] 1. Isolation, purification and culture of skin stem cells:

[0026] 1.1. In vitro separation of epidermal cells: soak and disinfect children’s foreskins after circumcision under sterile conditions with 0.05% chlorhexidine acetate for 10 minutes, rinse with normal saline, remove subcutaneous tissue, cut into 2mm×2mm size, place Digest in 0.25% trypsin / ethylenediaminetetraacetic acid solution at 37°C for 30 minutes to 1 hour; or place in 0.25% discrete enzyme II at 4°C for 24 hours to separate the epidermis and dermis; put the epidermis at 200 Grinding on a mesh sieve to collect the filtered cells;

[0027] 1.2. Purification: enrich the skin stem cells, spread the bottom of the culture dish with 50 μg / ml collagen IV solution, wait for drying, and divide the harvested epidermal cells into 1×10 5 / cm 2 Density planted on collagen IV, at 37°C for 10 minutes, suck off non-ad...

technical field

[0001] The invention relates to artificial skin, in particular to a method for preparing skin stem cell collagen sponge membrane artificial active skin. Background technique

[0002] Tissue-engineered artificial skin is the first relatively mature tissue engineering product to be developed for commercial application. Currently, its representative products are Biobrane TM , Integra TM 、Dermagraft TM and Apligraft TM They have played a certain role in the treatment of diseases such as burns, chronic venous ulcers and congenital skin deformities. However, to date, no product has been constructed using human skin stem cells as seed cells. Contents of the invention

[0003] The object of the present invention is to provide a method for preparing artificial active skin using human skin stem cells as seed cells for repairing, maintaining and improving skin tissue functions, simple preparation method and low cost skin stem cell collagen sponge membrane . [...

Examples

Embodiment Construction

[0024] The present invention is described in further detail below in conjunction with embodiment.

[0025] 1. Isolation, purification and culture of skin stem cells:

[0026] 1.1. In vitro separation of epidermal cells: soak and disinfect children’s foreskins after circumcision under sterile conditions with 0.05% chlorhexidine acetate for 10 minutes, rinse with normal saline, remove subcutaneous tissue, cut into 2mm×2mm size, place Digest in 0.25% trypsin / ethylenediaminetetraacetic acid solution at 37°C for 30 minutes to 1 hour; or place in 0.25% discrete enzyme II at 4°C for 24 hours to separate the epidermis and dermis; put the epidermis at 200 Grinding on a mesh sieve to collect the filtered cells;

[0027] 1.2. Purification: enrich the skin stem cells, spread the bottom of the culture dish with 50 μg / ml collagen IV solution, wait for drying, and divide the harvested epidermal cells into 1×10 5 / cm 2 Density planted on collagen IV, at 37°C for 10 minutes, suck off non-ad...