%0 Journal Article
%T Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures.
%A Boonekamp KE
%A Kretzschmar K
%A Wiener DJ
%A Asra P
%A Derakhshan S
%A Puschhof J
%A López-Iglesias C
%A Peters PJ
%A Basak O
%A Clevers H
%J Proc Natl Acad Sci U S A
%V 116
%N 29
%D 07 2019 16
%M 31253707
%F 12.779
%R 10.1073/pnas.1715272116
%X 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.