Skin regeneration is intricately controlled by epidermal stem cells. In human skin, the long-lived, slow-cycling, and highly proliferative stem cells are located in the basal layer of the interfollicular epidermis (IFE). The ability to isolate and culture human IFE stem cells (IFESCs) offers fascinating therapeutic potential for skin diseases as well as epithelial tissue engineering. Here we describe a straightforward strategy for generation of β1 integrin(+)/CD24(-) IFESCs from scalp with defined, serum-free, feeder-free medium and collagen I-coated culture plates. The use of defined media throughout isolation and cultivation allows for detailed investigation of the molecular events involved in ESC self-renewal and differentiation as well as provides a safe source for clinical use.
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