Hair follicle stem cells (HFSCs) from vibrissae follicles were isolated from the dermal papilla (DP) and dermal sheath (DS) of 3-month-old female Wistar rats. Cells have a fibroblast-like appearance and can be differentiated into adipocytes and osteocytes.
Dermal papilla (DP) and dermal sheath (DS) cells were dissected from vibrissae follicles and cultured for 5 days in minimal essential medium (MEM) supplemented with fetal bovine serum (FBS, 10%), DP or DS primary culture-conditioned medium (CM) and antibiotics. Cells were then dissociated in trypsin (0.25%) for 5 minutes, plated and cultured in MEM plus CM for 28 days with medium change every 7 days. Clones were further expanded for 16-35 days in MEM plus CM, with a medium change every 3-4 days. Clones were then transferred and maintained in MEM supplemented with FBS (10%), with a medium change every 3-4 days and were passaged every 21-35 days.