Construction method of 3D epidermis model
A construction method and epidermis technology, applied in 3D culture, artificial cell constructs, epidermal cells/skin cells, etc., can solve the problems of histological sectioning and result analysis, cumbersome support processing, and lack of theoretical support, etc., to achieve Facilitate attachment and growth, reduce exogenous added factors, and maintain metabolic activity
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Embodiment 1
[0033] The construction method of the 3D epidermis model of the present embodiment comprises the following steps:
[0034] Step 1. Resuscitate P2 generation human fibroblasts, perform routine culture on the revived fibroblasts, digest and passage after the cell plating rate reaches about 85%, and inoculate the culture medium containing serum at a density of 6×104 / cm2 after counting the cells After cultivating in a 37°C, 5% CO2 incubator for 24 hours, the culture solution was discarded, and then the culture solution containing mitomycin C was added, treated in a carbon dioxide incubator for 2 hours, and the culture solution was discarded. The hot PBS buffer solution gently washes the cells 3 times to obtain a fibroblast trophoblast; the serum-containing culture medium is based on a low-sugar DMEM culture medium, and fetal bovine serum is added, and the fetal bovine serum in the serum-containing culture medium is The volume percentage of serum is 10%; the culture fluid containin...
Embodiment 2
[0038] The construction method of the 3D epidermis model of the present embodiment comprises the following steps:
[0039] Step 1. Resuscitate the P2 generation human fibroblasts, perform routine culture on the revived fibroblasts, digest and passage after the cell plating rate reaches about 85%, and inoculate the cultured cells containing serum at a density of 3×104 / cm2 after counting the cells After cultivating in a 37°C, 5% CO2 incubator for 18 hours, the culture solution was discarded, and then the culture solution containing mitomycin C was added, treated in a carbon dioxide incubator for 3 hours, and the culture solution was discarded. Gently wash the cells with hot PBS buffer solution 5 times to obtain a fibroblast trophoblast; the serum-containing culture medium is based on α-MEM culture medium, and fetal bovine serum is added, and the fetal bovine serum in the serum-containing culture medium is The volume percentage of serum is 5%; the culture solution containing mito...
Embodiment 3
[0043] The construction method of the 3D epidermis model of the present embodiment comprises the following steps:
[0044] Step 1. Resuscitate P2 generation human fibroblasts, perform routine culture on the revived fibroblasts, digest and passage after the cell plating rate reaches about 85%, and inoculate the cultured cells containing serum at a density of 5×104 / cm2 after counting the cells After cultivating in a 37°C, 5% CO2 incubator for 20 hours, the culture solution was discarded, and then the culture solution containing mitomycin C was added, treated in a carbon dioxide incubator for 6 hours, and the culture solution was discarded. The hot PBS buffer solution gently washed the cells 4 times to obtain a fibroblast trophoblast; the serum-containing culture medium was based on DF12 culture medium, and fetal bovine serum was added, and the content of fetal bovine serum in the serum-containing culture medium was The volume percent content is 15%; the culture solution containi...
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