Hair follicle stem cells (HFSCs) were obtained from Wistar rat vibrissae follicles. Cells form new hair follicles, epidermis and sebaceous glands when transplanted to nude mice.
Vibrissae follicles were dissected and cultured for 3-7 days in William’s E medium supplemented with fetal bovine serum (FBS, 15%), penicillin and streptomycin, with medium change every 2 days. To obtain hair follicle stem cells (HFSCs), cells were incubated for 3-5 minutes in a solution of trypsin (0.1%) and Ethylenediaminetetraacetic acid (EDTA, 0.008%), to detach non-HFSCs. Adherent cells were washed with phosphate buffered saline (PBS) and dissociated in a solution of trypsin (0.1%) and Ethylenediaminetetraacetic acid (EDTA, 0.008%) for an additional 5 minutes. Cells were co-cultured with fibroblasts and passaged every 2 weeks.