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S. Selvam, S.C. Yiu, R.E. Smith, M.D. Trousdale, C.A. Peng, Z. Zhu, D. Stevenson, A.K. Mircheff, T. Nakamura, J.T. Jacob; Analysis of Rabbit Lacrimal Gland Acinar Cells cultured on ECMP Coated Silicone Sheets . Invest. Ophthalmol. Vis. Sci. 2005;46(13):4410.
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Purpose: To study the response of rabbit lacrimal acinar cells to extracellular matrix protein (ECMP) coated and uncoated silicone sheets. Methods: Purified rabbit lacrimal gland acinar cells were obtained from aseptically excised inferior lacrimal glands using standard protocols. Biomedical grade silicone sheets were cut, sterilized and placed in wells of a 24–well tissue culture plate. The silicone in half of the plate wells (12 wells) was then coated with matrigel, an ECMP containing preparation; the remaining 12 wells served as controls with no coating. Cell suspensions (5 x 105/ml) were seeded into all wells and the plate cultured at 37ºC under 5% CO2 for 5 days. On culture day 5, three wells of each type were fixed with Karnovsky's fixative for at least 2 h at 4oC and washed with PBS for scanning electron microscopy (SEM) to evaluate the cellular morphologic properties. Additionally, on the same cell culture day the cellular physiologic properties were examined. Four wells of each material type, two wells with carbachol stimulation (stimulated) and two without carbachol (unstimulated), were used to evaluate secretion of catalytically active beta hexosaminidase. Results: SEM pictures showed cell clusters (acini) on both the matrigel–coated and control groups. The surfaces of the cell clusters had the characteristic appearance of the basal side of acini with no microvilli. Neither material had a cellular monolayer present at the end of cell culture. A significant increase in cell secretory activity was found in the cells with carbachol stimulation. But the degree of stimulation between the two material groups was not statistically different. Conclusions: Silicone is highly biocompatible to purified rabbit lacrimal gland acinar cells. It supports and maintains the lacrimal acinar phenotype and shows promise for use in a tissue engineered tear secretory system.
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