关键词: adult stem cells interfollicular epidermis mammalian epidermis organoids

Mesh : Adult Stem Cells / physiology Animals Cell Culture Techniques / methods Cell Differentiation Cell Proliferation Cells, Cultured Epidermis / physiology Gene Knock-In Techniques Keratinocytes / physiology Mice Mice, Inbred C57BL Mice, Transgenic Organoids / physiology Receptors, G-Protein-Coupled / genetics Time Factors

来  源:   DOI:10.1073/pnas.1715272116   PDF(Sci-hub)   PDF(Pubmed)

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.
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