Hair follicle stem cells were isolated from mouse dorsal skin. Cells express various epithelial markers and have the capacity to differentiate into adipocytes, sebocytes and proteoglycan-producing cells.
Dorsal skin was removed and digested overnight, at 4°C, with dispase (2.5 mg/ml) in DMEM/F-12 medium.
The tissue was then subjected to a second digestion with
trypsin–EDTA (0.05%) for 5 min, washed, and plated onto tissue culture plates in DMEM/F12 (3:1) medium, supplemented with fetal calf serum (FCS, 10%), B27 supplement (20 μl/ml), basic fibroblast growth factor (bFGF, 20 ng/ml), and penicillin/streptomycin (1%).