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A. G. Jorge, A. C. Dias, L. Roma, C. M. Modulo, R. B. Filho, E. M. Rocha; Establishment of Lacrimal Gland Primary Acinar Cells’ Culture. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5329.
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Previous studies have shown that systemic diseases affect lacrimal gland function. Moreover, hormones and inflammatory mediators have been implicated in the pathogenesis; although their direct action over acinar cells function is not completely defined. To further understand the physiopathological steps of those diseases, our aim is to establish a lacrimal gland acinar cell culture line and to promote changes in culture conditions to evaluate the impact on secretory parameters.
Acinar cells of lacrimal gland of Wistar male rats were isolated and seed in culture plastic plates (1,7 x 106 cells, treated or not with fibronectin (5 µg/cm2 ), Matrigel (40 µg/ml), poly-L-lisine (PLL) (5 µg/cm2) and maintained with culture medium DMEM supplemented with 1 mM putrescine, 10 µg/ml reduced glutathione, 10µ/ml dexametasone, a mix of insulin-transferrin-sodium selenite (5 µg/ml: 5µg/ml: 5ng/ml), and 10 ng/ml EGF. The adherence and presence of peroxidase and insulin in the supernatant, with or without Carbachol (100µM) stimulation was evaluated along of a culture period of 14 days.
There was no adherence in plastic basis. Matrigel presented the higher level of cell adherence compared with PLL, plastic and fibronectin. In the mentioned conditions the supernatant levels of peroxidase or insulin was not affected by carbachol stimulation after 14 days.
Our work confirms that extra cellular matrix is a key element in maintenance of lacrimal gland cells in culture. Further studies are necessary to correlate culture conditions and time course of biomarkers expression in order to follow lacrimal gland acinar cells proliferation, maintenance and function in culture.
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