Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures.
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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.
Boonekamp, K. E., Kretzschmar, K., Wiener, D. J., Asra, P., Derakhshan, S., Puschhof, J., ... Clevers, H.
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Boonekamp, Kim E||Kretzschmar, Kai||Wiener, Dominique J||Asra, Priyanca||Derakhshan, Sepideh||Puschhof, Jens||López-Iglesias, Carmen||Peters, Peter J||Basak, Onur||Clevers, Hans