Abstract
A novel lingual epithelial organoid culture system using a three-dimensional (3D) matrix and growth factors has recently been established. In the culture system, organoids with multilayered squamous keratinized epithelium and typical morphological features of filiform papillae are generated from single lingual epithelial cells at a high efficiency. The culture system is created in order to observe the differentiation and maturation process of each lingual epithelial stem cell and to observe abnormal organoid formation from malignant cells obtained from carcinogen-treated mice. Thus, our culture system will contribute to the advancement of research into the regulatory mechanism of lingual epithelium and the underlying mechanisms of carcinogenesis.
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References
Hume WJ (1986) Kinetics of cell replacement in the stratum granulosum of mouse tongue epithelium. Cell Tissue Kinet 19:195–203
Li F, Cao J, Zhou M (2012) NTPDase 2+ cells generate lingual epithelia and papillae. Front Genet 3:255. doi:https://doi.org/10.3389/fgene.2012.00255
Howard JM, Nuguid JM, Ngole D, Nguyen H (2014) Tcf3 expression marks both stem and progenitor cells in multiple epithelia. Development 141:3143–3152
Tanaka T, Komai Y, Tokuyama Y, Yanai H, Ohe S, Okazaki K, Ueno H (2013) Identification of stem cells that maintain and regenerate lingual keratinized epithelial cells. Nat Cell Biol 15:511–518
Mbiene JP, Maccallum DK, Mistretta CM (1997) Organ cultures of embryonic rat tongue support tongue and gustatory papilla morphogenesis in vitro without intact sensory ganglia. J Comp Neurol 377:324–340
Zhou Y, Liu HX, Mistretta CM (2006) Bone morphogenic proteins and noggin: inhibiting and inducing fungiform taste papilla development. Dev Biol 297:198–213
Ookura T, Kawamoto K, Tsuzaki H, Mikami Y, Ito Y, Oh SH, Hino A (2002) Fibroblast and epidermal growth factors modulate proliferation and neural cell adhesion molecule expression in epithelial cells derived from the adult mouse tongue. In Vitro Cell Dev Biol Anim 38:365–372
Luo X, Okubo T, Randell S, Hogan BLM (2009) Culture of endodermal stem/progenitor cells of the mouse tongue. In Vitro Cell Dev Biol Anim 45:44–54
Hisha H, Tanaka T, Kanno S, Tokuyama Y, Komai Y, Ohe S, Yanai H, Omachi T, Ueno H (2013) Establishment of a novel lingual organoid culture system: generation of organoids having mature keratinized epithelium from adult epithelial stem cells. Sci Rep 3:3224. doi:https://doi.org/10.1038/srep03224
Flint N, Cove FL, Evans GS (1991) A low-temperature method for the isolation of small-intestinal epithelium along the crypt-villus axis. Biochem J 280:331–334
Red-Horse K, Ueno H, Weissman IL, Krasnow M (2010) Coronary arteries form by developmental reprogramming of venous cells. Nature 464:549–553
Rinkevich Y, Lindau P, Ueno H, Longaker MT, Weissman IL (2011) Germ-layer and lineage-restricted stem/progenitors regenerate the mouse digit tip. Nature 476:409–413
Tang X-H, Knudsen B, Bemis D, Tickoo S, Gudas LJ (2004) Oral cavity and esophageal carcinogenesis modeled in carcinogen-treated mice. Clin Cancer Res 10:301–313
Acknowledgement
This work was financially supported by the Funding Program for Next Generation World-Leading Researchers, The Mochida Memorial Foundation, The Naito Memorial Foundation, The Cell Science Research Foundation, The Uehara Memorial Foundation, The Mitsubishi Foundation, and The Yasuda Memorial Foundation to H.U., and a grant from a Grant-in-aid for Scientific Research (C) 23590953 to H.H.
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Hisha, H., Ueno, H. (2016). Organoid Culture of Lingual Epithelial Cells in a Three-Dimensional Matrix. In: Turksen, K. (eds) Organoids. Methods in Molecular Biology, vol 1576. Humana, New York, NY. https://doi.org/10.1007/7651_2016_3
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DOI: https://doi.org/10.1007/7651_2016_3
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