Characterization Of Murine Lacrimal Gland Epithelial Cell Line

Abstract

Purpose: : Regeneration of lacrimal gland is necessary to reconstruct severely damaged ocular surface with no wetness. To analyze the function of lacrimal gland in cellular level, lacrimal gland cell line might be useful. Because of no report and no availability of such cell lines, we tried to establish the murine lacrimal gland cell lines spontaneously.

Methods: : Exorbital lacrimal glands of 3-week old mice (C57B/6) were dissected. The tissues were treated with Collagenase/Hyarulonidase for digestion. Cells were seeded on plastic dishes in culture medium (Cnt07 medium with cholerae toxin). Primary culture were passaged at the time of semi-confluent. RNA was extracted for real time PCR of E-cadherin and Lactoferrin. Immunostaining of antibodies against Cytokeratin 14, Aquaporin 8 and alpha-SMA was performed. At the time of passage 20, limited dilution of the cells were performed for cloning of the cells.

Results: : Long term culture of murine lacrimal gland epithelial cells was established spontaneously in Cnt07 culture medium with cholerae toxin. After clomg by limiting dilution, cell were maintained more than 1 year, which showed E-cadherin positive, cytokeratin 14 positive, alpha-SMA negative phenotype confirmed by immunostaining and real time PCR.

Conclusions: : Murine lacrimal gland epithelial cells line was established spontaneously, and coule be used for future research.

Keywords: lacrimal gland • immunohistochemistry