Tail skin was removed from the tail bone and incubated overnight at 4°C in Hanks’ balanced salt solution (HBSS) (Gibco) and 0.25% trypsin (Gibco). The epidermis was then separated from the dermis and incubated on a rocking plate (100 rpm) at room temperature for 5 min. Basal cells were mechanically separated from the epidermis by flushing 10 times under the epidermis. Trypsin was then neutralized by adding Dulbecco’s modified Eagle’s medium. (DMEM) (Gibco) supplemented with 5% Chelex fetal calf serum (FCS) and filtrated on a 70-μm filter (Falcon). Cells were cultured in MEM supplemented with 10% FBS, hydrocortisone (0.4 μg/ml), epidermal growth factor (10 ng/ml), 2 × 10−9 MT3, 1% penicillin/streptomycin, 2 mM l-glutamine and incubated at 37°C with 20% O2 and 5% CO2. Twenty-four hours later, the cells were treated with recombinant vascular endothelial growth factor (VEGF) (50 ng/ml; R&D).

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