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M. Alves, A. C. Dias, C. M. Modulo, L. Roma, A. G. Jorge, L. T. Malki, E. M. Rocha; The Role of the Extracellular Matrix in the Function and Morphology of Lacrimal Gland Primary Acinar Cell Culture. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4237.
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The relationship between lacrimal gland acinar cells and the extracellular matrix influences the cell phenotype, metabolism and replacement. To further understand the physiopathological aspects of this interaction, the aim of this study was to promote changes in primary lacrimal gland acinar cell culture conditions to evaluate their impact on cell morphology and secretory parameters.
Acinar cells of the lacrimal gland of male Wistar rats were isolated and seeded on plastic culture plates, treated or not with Matrigel (40 µg/ml), poly-L-lysine (PLL) (5 µg/cm2), or amniotic membrane and cultured with DMEM supplemented with 1 mM putrescine, 10 µg/ml reduced glutathione, 10 µ/ml dexamethasone, a mix of insulin-transferrin-sodium selenite (5 µg/ml: 5µg/ml: 5ng/ml), 10 ng/ml EGF, and 10% FBS. The adherence, viability, cell morphology and presence of peroxidase in the supernatant, with or without carbachol (100 µM) stimulation were evaluated over a culture period of 11 days.
Cell size, adherence and aggregation were superior in Matrigel substrate compared to PLL and amniotic membrane. There was no adherence to the plastic base. The cell number and viability increased until day 7 and declined thereafter in matrigel and PLL substrate. Under the mentioned conditions, peroxidase levels in the supernatant were highly affected by carbachol stimulation in the matrigel group.
Our work confirms that the extracellular matrix is a key element in the maintenance of lacrimal gland acinar cells in culture. Moreover, it indicates that acinar cells maintain the physiological response of secretion for an initial period of time.Financial Support: FAPESP, FAEPA, CNPq, CAPES
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