Stem cells were isolated from hair follicle-containing, full-thickness scalp skin samples. Cells express mesenchymal markers and have osteogenic, chondrogenic and adipogenic differentiation capacities.
Skin tissue was washed extensively and trimmed to remove the fat tissue layer. Then, it was cut into small pieces and digested in collagenase type I (0.5%) for 4 hours. After digestion, hair follicles were isolated using forceps, filtered through a 40 μm cell strainer, and washed several times with phospho-buffered saline (PBS). Hair follicles were then transferred to a 96-well plate and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with fetal bovine serum (FBS;10%) to allow for migration of cells to the plate. Cells that migrated from the dermal sheath or papilla and had the morphological appearance of mesenchymal cells were isolated and expanded.