Background– Nestin, which was originally described as a neural crest stem cell marker, is known to be expressed in bulge follicle cells of human, canine and murine anagen hairs. However, the capacity of nestin-expressing cells to differentiate into the components of the hair follicle or the epidermis has been insufficiently investigated. Hypothesis/objectives – To determine whether nestin-expressing cells are capable of differentiating into keratinocytes. Conclusions and clinical importance – Our results suggest that progenitor cells that differentiate into ORS keratinocytes are distinct from those for other hair follicle or epidermal components. These findings provide implications for regenerative medicine and the molecular classification of hair follicle tumours.