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PriCells: Primary dermal fibroblasts isolated from human
PriCells: Primary dermal fibroblasts isolated from human
Fibroblasts were established from donors of human normal skin, scar, and keloid using the explant method.
The specimens were washed with sterile phosphate-buffered saline (PBS), and the subcutaneous tissues were removed carefully.
The skins were then cut into small pieces (1-2 mm3).
After being digested with PriCells Isolation Kit III (or using 0.25% trypsin containing 0.05% ethylenediaminetetraacetic acid) at 4°C overnight, the epidermal layers were removed, and the remaining dermal parts were further digested with PriCells Isolation Kit III (or using 0.25% trypsin containing 0.05% ethylenediaminetetraacetic acid) at 37°C for another 4 hours.
The digested cells were then passed through a 70-µm cell strainer (BD Biosciences), centrifuged, and resuspended in PriCells primary cell culture system.
Cells were seeded on tissue culture plates at 1x103 cells/cm2, and maintained at 37°C with 5% CO2.
After 24 hours, the plates were washed with PBS to remove residual non-adherent cells.
The resulting adherent cells were grown to confluency within 7 days.
Cells were subcultured every 4 days using 0.25% trypsin/EDTA.