Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures
Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures
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DOI：
10.1073/pnas.1715272116
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发表时间：
2019-07-16
影响因子：
11.1
通讯作者：
Clevers, Hans
中科院分区：
文献类型：
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作者：
Boonekamp, Kim E.;Kretzschmar, Kai;Clevers, Hans
哺乳动物表皮干细胞维持皮肤表皮的稳态，并有助于其在整个成年期的再生。虽然二维小鼠表皮干细胞培养在几十年前就建立起来了，但一个长期的、饲养细胞和无血清的培养系统还没有重现小鼠表皮结构。在这里，我们描述了一个表皮类器官培养系统，允许成人表皮干细胞的长期，遗传稳定的扩张。我们的表皮扩张培养基结合了非典型的高钙浓度、cAMP、FGF和R-spondin信号的激活以及骨形态发生蛋白（BMP）信号的抑制。类器官从成年小鼠皮肤中建立，并在至少6个月的时间内扩张，同时保持小鼠滤泡间表皮的基底-顶端组织。该系统是体外研究表皮稳态和疾病的有力工具。
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.