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Objective To differentiate urine derived pluripotent stem cells (iPSCs) into hair follicle stem cells (HFSCs) to construct a skin substitutes. Methods Human urinary cells isolated from a healthy donor were re-programmed into iPSCs. iPSCs were then induced into iPS-HFSCs. Immunocytochemistry and RT-qPCR were used to examine the characteristics of the induced iPS-HFSCs. Results The iPSCs induced from urine cells have characteristics as embryonic stem cells. iPSCs derived cells displayed the morphology, genes expression and phenotypes similar to iPS-HFSCs. Conclusions The urine cells are successfully reprogramed into iPSCs by four transcription factors, and iPSCs derived cells show characteristics similar to HFSCs.