A method for preparing composite film for treating vitiligo by using autologous hair follicle melanocytes
A melanocyte and composite membrane technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of scars that are not suitable for young children and children, high treatment costs, limited skin lesions, etc., and achieve a good cell phase. Capacitance, promoting adhesion and proliferation, enhancing effect
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Embodiment 1
[0028] (1) Preparation of collagen film: Prepare a collagen solution with a concentration of 5 wt% with an acetic acid solution with a concentration of 1 wt%, stir well, spread it flat and dry to form a film to obtain a pure collagen film ( Figure 4 Middle A).
[0029] (2) Preparation of autologous hair follicle melanocytes: Use a 2mm skin puncher to take 2 scalps containing hair follicles, soak them in 70% alcohol, rinse with PBS solution, rinse with sterile saline until the surface is clean, and immediately add penicillin (400U / mL), streptomycin (400mg / mL) and Tylosin (250mg / mL) in PBS solution for 3 times, then immersed in sterile 0.25% neutral protease, digested at 4°C for 5h and put into new PBS solution , take out the hair follicles under a microscope and place them in the medium, slowly add 0.25% trypsin dropwise, incubate at 37°C for 10 minutes, remove excess trypsin, replace with new medium, 37°C, 5% CO 2 Cell separation can be observed by culturing in a saturated ...
Embodiment 2
[0032] (1) Preparation of collagen film: Prepare a collagen solution with a concentration of 5 wt% with acetic acid solution with a concentration of 1 wt%, stir well, lay flat and dry to form a film to obtain a pure collagen film.
[0033] (2) Preparation of autologous hair follicle melanocytes: same as in Example 1. For the repair area greater than 20cm 2 In the affected area, the cells expanded in vitro were completed for clinical use within 2 weeks.
[0034] (3) Transfer the cultured autologous melanocytes to the surface of the collagen membrane, and then give it to patients with vitiligo. When using it, you only need to simply grind the treatment area, and then paste the autologous melanocytes on the treatment area to complete the treatment; after 4 weeks, you can see obvious recoloration.
Embodiment 3
[0036] (1) Preparation of collagen / PVA composite film: prepare a collagen solution with a concentration of 0.5wt% with an acetic acid solution with a concentration of 0.3wt%, and set aside; weigh PVA, add it to deionized water, and prepare a PVA solution with a concentration of 5wt% , stirred in a constant temperature oil bath at 65°C for 6 hours, added collagen solution after dissolving, prepared a mixed solution with a volume ratio of PVA and collagen of 1:10, stirred well, laid flat and dried to form a film to obtain a collagen / PVA composite film ( Figure 4 Middle B).
[0037] (2) Preparation of autologous hair follicle melanocytes: same as in Example 1. For the repair area less than 20cm 2 In the affected area, the cells expanded in vitro are ready for clinical use within 1 week.
[0038] (3) Transfer the cultured autologous melanocytes to the surface of the collagen / PVA composite membrane, and then give it to patients with vitiligo. When using it, you only need to sim...
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