Human Dermal Fibroblast Extraction and Culture

The lines of human dermal fibroblasts were obtained from donor skin, as described previously [10 (link),13 (link),14 (link)]. Briefly, tissue biopsies were collected from patients (who have given the informed contest) during surgery. The study was approved by the Ethics Committee of the Institute of Medical Cell Technologies, Ekaterinburg. Skin biopsies were cut into small pieces, and cells were extracted by tissue dissociation method. Extracted fibroblasts were grown in culture flasks (Nunc, Roskilde, Denmark) at 37 °C and 5% CO₂. Cells were passaged when they covered 80% of the flask surface area. Then sub-culturing cells were treated with 0.25% trypsin-EDTA solution (Gibco, Thermo Fisher Scientific, Inc., Waltham, MA, USA). Cell number was estimated by cell counter TC-20 device (Bio-rad, Hercules, CA, USA). The cell viability was measured by staining with trypan blue. Fibroblasts were stored in liquid nitrogen.