Application of ALDH3A1/IL-17 axis in preparation of medicine for treating refractory ulcer
A technology for refractory ulcers and drugs, applied in the field of biomedicine, can solve problems such as carcinogenicity, easy degradation, and unbearable for patients, and achieve the effect of reducing inflammatory infiltration and enhancing cell proliferation
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[0048] 1. Methods and Materials
[0049] 1. Cell culture and transfection
[0050] The human keratinocyte cell line HaCaT was purchased from Cell Lines Service, Eppelheim, Germany. Cells were cultured in DMEM-high glucose (4.5 g / L, Gibco, Life Technologies, Carlsbad, CA) containing 10% (v / v) fetal bovine serum (HyClone, Logan, UT, USA) and 1% ( v / v) Streptomycin-penicillin solution (10000 U / mL, Gibco, Life Technologies) with or without 20 ng / ml bFGF (basic fibroblast growth factor, ThermoFisher Science, #13256029) at 37°C Constant temperature culture.
[0051] ALDH3A1-overexpressed HaCaT cells were constructed using a lentiviral transfection system. Using DNAFect transfection reagent (109120002, HarOLife, Shanghai, China), 1.7 μg ALDH3A1 overexpression vector (backbone plasmid is pLVX-IRES-Puro (Catalog No.632183, Clontech, US), see figure 1 , the ALDH3A1 sequence directional cloned into the vector backbone is shown in SEQ ID NO: 7, the insertion site is EcoRI / NotI) or CMV...
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