Method for constructing skin tissue
A construction method and skin technology, applied in the field of cell culture, can solve the problems of expensive patients and restrictions on the development and application of tissue engineered skin, and achieve the effect of convenient control
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Embodiment 1
[0070] A kind of keratinocyte and fibroblast, adopt following method to prepare:
[0071] Configure the following media:
[0072] Optimized Green culture solution: DMEM medium and Ham's F12 nutrient solution were mixed at a volume ratio of 3:1, and then the following components were added based on the volume after the two were mixed: FCS with a volume percentage of 10%, EGF 10ng / ml, hydrocortisone 0.4μg / ml, adenine 0.18mM, insulin 5μg / ml, transferrin 5μg / ml, glutamine 2mM, iodothyronine 0.2μM, amphotericin B 0.625μg / ml, Penicillin 100U / ml, streptomycin 100μg / ml.
[0073] 1. Skin Biopsy Sampling
[0074] 1.1 Under sterile conditions, take 1.5cm of inner thigh skin tissue 2 The size was placed in DMEM containing 10% newborn calf serum (FBS) and double antibodies (100 IU / ml penicillin, 100 ug / ml streptomycin), and stored at 4°C.
[0075] 1.2 The skin tissue was cut into three pieces of 1x5 mm in size, and washed 10 times with phosphate-buffered saline (PBS) containing 50 U / m...
Embodiment 2
[0099] A method for constructing skin tissue, comprising the steps of:
[0100] Configure the following media:
[0101] Optimized Green culture solution: DMEM medium and Ham's F12 nutrient solution were mixed at a volume ratio of 3:1, and then the following components were added based on the volume after the two were mixed: FCS with a volume percentage of 10%, EGF 10ng / ml, hydrocortisone 0.4μg / ml, adenine 0.18mM, insulin 5μg / ml, transferrin 5μg / ml, glutamine 2mM, iodothyronine 0.2μM, amphotericin B 0.625μg / ml, Penicillin 100U / ml, streptomycin 100μg / ml.
[0102] The fibroblasts and keratinocytes prepared in Example 1 were used.
[0103] Seed fibroblasts in scaffolds at a density of 5x10 5 / cm 2 , add 1ml of optimized Green culture medium to the scaffold.
[0104] Add 2ml of RPMI1640 culture solution around the scaffold.
[0105] 37°C, 5% CO 2 Cultured for 3 days, changing the culture medium every 2 days.
[0106] On the fourth day, keratinocytes (1x10 6 cells / ml), adh...
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