@article{9d1e3e4bc97b4953a54a4926280b9ea5,
title = "Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures",
abstract = "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.",
keywords = "mammalian epidermis, organoids, adult stem cells, interfollicular epidermis, HAIR FOLLICLE, SKIN, KERATINOCYTES, PROLIFERATION, DESMOPLAKIN, GROWTH, CLASSIFICATION, ORGANIZATION, HOMEOSTASIS, PLASTICITY",
author = "Boonekamp, {Kim E.} and Kai Kretzschmar and Wiener, {Dominique J.} and Priyanca Asra and Sepideh Derakhshan and Jens Puschhof and Carmen Lopez-Iglesias and Peters, {Peter J.} and Onur Basak and Hans Clevers",
note = "Funding Information: ACKNOWLEDGMENTS. We thank Joep Beumer for advice and critical reading of the manuscript, as well as contributions in the early stages of the project; Wilma Hoevenaar for imaging of the H2B-mNeonGreen–transduced organoids; Maaike van den Born and Johan H. van Es for help with animal procedures; Reinier van den Linden for and Stefan van der Elst help with flow cytometry; the Utrecht Sequencing Facility (USEQ) for sequencing; Anja van de Stolpe and Philips Research for their contributions to the project; the technical staff of the Microscopy CORE Lab of M4I Nanoscopy in Maastricht University and Willine van de Wetering for all their support in microscopy; and Anko de Graaff and the Hubrecht Imaging Center for help with imaging. H.C. gratefully acknowledges financial support from the Swiss National Science Foundation (Sinergia Grant CRSII3 160738-1) and the Netherlands Organization for Scientific Research (NWO-ZonMW, 114021012). This work is part of the Oncode Institute, which is partly financed by the Dutch Cancer Society and was funded by the gravitation program CancerGenomiCs.nl from the Netherlands Organization for Scientific Research (NWO). K.K. is a long-term fellow of the Human Frontier Science Program Organization (LT771/2015) and was the recipient of a VENI grant (NWO-ZonMW, 016.166.140). Funding Information: We thank Joep Beumer for advice and critical reading of the manuscript, as well as contributions in the early stages of the project; Wilma Hoevenaar for imaging of the H2B-mNeonGreen–transduced organoids; Maaike van den Born and Johan H. van Es for help with animal procedures; Reinier van den Linden for and Stefan van der Elst help with flow cytometry; the Utrecht Sequencing Facility (USEQ) for sequencing; Anja van de Stolpe and Philips Research for their contributions to the project; the technical staff of the Microscopy CORE Lab of M4I Nanoscopy in Maastricht University and Willine van de Wetering for all their support in microscopy; and Anko de Graaff and the Hubrecht Imaging Center for help with imaging. H.C. gratefully acknowledges financial support from the Swiss National Science Foundation (Sinergia Grant CRSII3 160738-1) and the Netherlands Organization for Scientific Research (NWO-ZonMW, 114021012). This work is part of the Oncode Institute, which is partly financed by the Dutch Cancer Society and was funded by the gravitation program CancerGenomiCs.nl from the Netherlands Organization for Scientific Research (NWO). K.K. is a long-term fellow of the Human Frontier Science Program Organization (LT771/2015) and was the recipient of a VENI grant (NWO-ZonMW, 016.166.140). Publisher Copyright: {\textcopyright} 2019 National Academy of Sciences. All rights reserved.",
year = "2019",
month = jul,
day = "16",
doi = "10.1073/pnas.1715272116",
language = "English",
volume = "116",
pages = "14630--14638",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "29",
}