Characterization of cultivated murine lacrimal gland epithelial cells

Shinya Kobayashi, Tetsuya Kawakita, Motoko Kawashima, Naoko Okada, Kenji Mishima, Ichiro Saito, Masataka Ito, Shigeto Shimmura, Kazuo Tsubota

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Purpose: To date, mouse lacrimal gland epithelial cells have been cultured successfully but only in cases involving newborn mouse lacrimal glands. In this work, we attempted to cultivate and characterize adult mouse lacrimal gland epithelial cells. Methods: Lacrimal glands were removed from newborn mice (C57B/6) and isolated lacrimal gland epithelial cells were seeded onto tissue culture treated or low adherent culture dishes in Cnt-07 culture medium with or without cholera toxin. Cultivated cells were characterized by immunostaining with pan-cytokeratin, α-smooth muscle actin, and lactoferrin antibodies. Lacrimal gland cells from 7-week-old green fluorescent protein (GFP) and non-GFP (C57B/6) mice were mixed and seeded onto uncoated dishes to assess sphere-forming efficiency. Cells were also seeded onto 3T3 cell feeder layers to assess colony forming efficiency. Results: Lacrimal gland epithelial cells were selectively cultured with cholera toxin, and cell type was verified by pancytokeratin and α-smooth muscle actin immunostaining. Sphere formation from single cells of adult mice was observed using specific medium and low adherent culture dishes. These cells could also undergo colony formation on 3T3 feeder cells. Conclusions: Adult mouse lacrimal gland epithelial cells were successfully cultivated in cholera toxin-containing medium, and were observed to form spheres from single cells.

Original languageEnglish
Pages (from-to)1271-1277
Number of pages7
JournalMolecular vision
Volume18
Publication statusPublished - 2012 May 12

ASJC Scopus subject areas

  • Ophthalmology

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  • Cite this

    Kobayashi, S., Kawakita, T., Kawashima, M., Okada, N., Mishima, K., Saito, I., Ito, M., Shimmura, S., & Tsubota, K. (2012). Characterization of cultivated murine lacrimal gland epithelial cells. Molecular vision, 18, 1271-1277.