Esophageal tissue constructed by tissue engineering
A tissue engineering, esophagus technology, applied in the field of biomedical engineering, can solve problems such as slow progress
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[0012] 1) Isolation and culture of epidermal keratinocytes
[0013] The epidermis of the patient was trimmed into strips with a width of 1-2mm, rinsed twice in PBS without calcium and magnesium, and immersed in 0.5% neutral protease overnight at 4°C with the epidermis facing upward. After the epidermis was torn off from the dermis, it was cut into pieces and digested with 0.05% trypsin-0.53mM EDTA at 37°C for 20 minutes, and 10ml of trypsin inhibitor was used to stop the digestion. Filter through a 150-mesh stainless steel filter, centrifuge (1200 rpm, 10 minutes), and count to make a single-cell suspension, press 3×10 6 / 75cm 2 Density seeding culture, serum-free epidermal cell medium (GIBRO, USA), plus bovine pituitary extract and recombinant epidermal growth factor, at 37 ° C, 5 % CO 2 , cultured in an incubator with saturated humidity, and the culture medium was changed every 3 days. When the cells reached 60-75% confluence, they were digested with 0.05% trypsin at 37°C...
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