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Significance A vast number of skin pathologies cannot be studied well in vitro because of the lack of a long-term feeder and serum-free culture system for epidermis. Here we develop a long-term culture system for adult murine keratinocytes that is genetically stable over time. Using an organoid-based approach, we establish a 3D culture of murine keratinocytes that resembles the interfollicular epidermis, including proliferative cells in the basal layer and differentiation toward the cornified envelope. As culture models of spontaneously differentiating murine epidermis, these organoids may therefore serve as a platform for studying the basic biology of skin diseases, such as cancer, and their underlying genetic alterations.

Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures