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Masataka Ito, Yoko Karasawa, Kouzo Harimoto, Nariyoshi Shinomiya, Junko Imaki, Masaru Takeuchi; Growth Factor-dependent Proliferation of Cultured Lacrimal Gland Epithelial Cells Derived from Embryonic and Newborn Mice. Invest. Ophthalmol. Vis. Sci. 2012;53(14):630.
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© ARVO (1962-2015); The Authors (2016-present)
Development of mouse lacrimal gland begins as a single budding from conjunctival epithelium near the temporal fornix at embryonic day 13.5 (E13.5). Fibroblast growth factor 10 (FGF10) secreted from sub-epithelial fibroblasts is known to be the inducer of the initial development of the gland. However, other growth factors involved in the development of this gland especially in the later stages are unknown. The aim of the present study was to analyze if epithelial growth factor (EGF) and hepatocyte growth factor (HGF) as well as FGF10 were involved in the development of lacrimal gland in the later embryonic and newborn stages of mice, in vitro.
Lacrimal gland epithelial cells isolated from E16-17 embryos and 0-2 day-old mice were cultured for three days in a fully-supplemented defined medium. Then the cells received a fresh supplement-free medium with a growth factor (FGF10, EGF, or HGF) or their receptor kinase inhibitors (SU5402, AG1478, or PHA665752, respectively) added alone or in combination to the medium. After three days, cell viability was measured by an assay of mitochondrial dehydrogenase activities and total cell death was measured by activities of lactate dehydrogenase released from dead cells to the medium.
Neither FGF10 nor SU5402 induced significant changes on cell viability or cell death. Administration of EGF or HGF raised cell viability while AG1478 and PHA665752 decreased cell viability and increased incidence of cell death. When the receptor inhibitor, AG1478 or PHA665752, was added together with its correspondent growth factor, the levels of cell viability was the same as the culture administrated the receptor inhibitor alone. Even in the presence of the receptor inhibitor, AG1478 or PHA665752, the irrespective growth factor, HGF or EGF, could improve cell viability but more weakly than the culture without the inhibitor. These results were observed in both cultures of the cells from embryonic and newborn mice.
In mouse lacrimal gland, growth factors involved in the epithelial proliferation were considered to change along with the development of the gland. In the late-embryonic and neonatal stages, proliferation of the epithelial cells depend more on EGF and HGF which act in an autocrine manner than FGF10, which was important in early development of the gland.
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