Hair follicle stem cells (HFSCs) were isolated from adult (50-65 year-old donors) or fetal (22–24 weeks) scalp tissues. Cells have multiple characteristics that are similar to neural crest cells (NCC), including expression of the NCC markers SNAI2 and SOX10. A fraction of the cells also express the embryonic stem cell transcription factors NANOG and POU5F1 (Oct-4). Microarray analysis showed that cells have a highly similar gene expression pattern to mouse epiblast neural crest stem cells (mEPI-NCSC). HFSCs are capable of self renewing, through asymmetric cell division, retain Bromodeoxyuridine (BrdU) label and proliferate as spheres. In addition, the cells can be differentiated into myogenic, melanocytic, mesenchymal and neuronal cell lineages.
Hair follicle stem cells (HFSCs) were isolated from hair follicles of adult (50-65 year-old donors) or fetal (22–24 weeks) scalp tissues. Follicle cells were dissociated for 30 minutes in trypsin (0.25%) containing ethylene diamine tetraacetic acid (EDTA, 0.03%) and filtered through a 40 um cell strainer. Cells were cultured in non-coated flasks in human embryonic stem cell (hESC) medium containing knockout Dulbecco’s modified Eagle’s medium/Nutrient Mixture F-12 (DMEM/F-12, 80%), knockout serum replacer (20%), L-glutamine (200 mmol/L), 2-mercaptoethanol (0.1 mmol/L), nonessential amino acids (1%) and basic fibroblast growth factor (bFGF, 4 ng/ml). Cells were maintained in conditioned hESC medium (hESC medium conditioned by mouse embryonic fibroblasts (MEFs)), mixed with hESC medium at a 3:1 ratio.