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

Boonekamp, Kim E; Kretzschmar, Kai; Wiener, Dominique J.; Asra, Priyanca; Derakhshan, Sepideh; Puschhof, Jens; López-Iglesias, Carmen; Peters, Peter J; Basak, Onur; Clevers, Hans (2019). Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures. Proceedings of the National Academy of Sciences of the United States of America - PNAS, 116(29), pp. 14630-14638. National Academy of Sciences NAS 10.1073/pnas.1715272116

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Mammalian epidermal stem cells maintain homeostasis of the skin epidermis and contribute to its regeneration throughout adult life. While 2D mouse epidermal stem cell cultures have been established decades ago, a long-term, feeder cell- and serum-free culture system recapitulating murine epidermal architecture has not been available. Here we describe an epidermal organoid culture system that allows long-term, genetically stable expansion of adult epidermal stem cells. Our epidermal expansion media combines atypically high calcium concentrations, activation of cAMP, FGF, and R-spondin signaling with inhibition of bone morphogenetic protein (BMP) signaling. Organoids are established robustly from adult mouse skin and expand over at least 6 mo, while maintaining the basal-apical organization of the mouse interfollicular epidermis. The system represents a powerful tool to study epidermal homeostasis and disease in vitro.

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