title: Long-term homeostasis and wound healing in an in vitro epithelial stem cell niche model.
abstract: Cultures of epithelial cells are limited by the proliferative capacity of primary cells and cell senescence. Herein we show that primary human epithelial cell sheets cultured without dermal equivalents maintained homeostasis in vitro for at least 1 year. Transparency of these sheets enabled live observation of pigmented melanocytes and Fluorescent Ubiquitination-based Cell Cycle Indicator (FUCCI) labeled epithelial cells during wound healing. Cell turn over and KRT15 expression pattern stabilized within 3 months, when KRT15 bright clusters often associated with niche-like melanocytes became apparent. EdU labels were retained in a subset of epithelial cells and melanocytes after 6 months chasing, suggesting their slow cell cycling property. FUCCI-labeling demonstrated robust cell migration and proliferation following wounding. Transparency and long-term (1 year) homeostasis of this model will be a powerful tool for the study of wound healing and cell linage tracing.
authors.affiliation: Department of ophthalmology/Corneal cell biology group, Keio university school of medicine, Tokyo, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Miyashita
authors.affiliation: Department of ophthalmology/Corneal cell biology group, Keio university school of medicine, Tokyo, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Niwano
authors.affiliation: Department of ophthalmology/Corneal cell biology group, Keio university school of medicine, Tokyo, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Yoshida
authors.affiliation: Department of ophthalmology/Corneal cell biology group, Keio university school of medicine, Tokyo, Japan.
authors.forename: Shin
authors.initials: S
authors.lastname: Hatou
authors.affiliation: Department of ophthalmology/Corneal cell biology group, Keio university school of medicine, Tokyo, Japan.
authors.forename: Emi
authors.initials: E
authors.lastname: Inagaki
authors.affiliation: Department of ophthalmology/Corneal cell biology group, Keio university school of medicine, Tokyo, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Tsubota
authors.affiliation: Department of ophthalmology/Corneal cell biology group, Keio university school of medicine, Tokyo, Japan.
authors.forename: Shigeto
authors.initials: S
authors.lastname: Shimmura
mesh_terms: D015415:Biomarkers; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000166:cytology; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006706:Homeostasis; D006801:Humans; D053569:Keratin-15 / Q000378:metabolism; D008544:Melanocytes / Q000378:metabolism; D055153:Stem Cell Niche; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014945:Wound Healing
chemical_list: D015415:Biomarkers; C116015:KRT15 protein, human; D053569:Keratin-15