Hair follicle stem cells (Chinese Academy of Sciences)

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.