Long-term homeostasis and wound healing in an in vitro epithelial stem cell niche model

Hideyuki Miyashita, Hiroko Niwano, Satoru Yoshida, Shin Hatou, Emi Inagaki, Kazuo Tsubota, Shigeto Shimmura

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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.

Original languageEnglish
Article number43557
JournalScientific Reports
Volume7
DOIs
Publication statusPublished - 2017 Feb 24

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