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Abstract #46436 Published in IGR 13-3
In vitro models for glaucoma research: effects of hydrostatic pressure
Lei Y; Rajabi S; Pedrigi RM; Overby DR; Read AT; Ethier CRInvestigative Ophthalmology and Visual Science 2011; 52: 6329-6339
PURPOSE: The response of cells (e.g., optic nerve head [ONH] cells) to mechanical stress is important in glaucoma. Studies have reported the biological effects of hydrostatic pressure on ONH cells cultured on a rigid substrate. An apparatus, designed to independently vary hydrostatic pressure and gas tension (including oxygen tension) in culture medium, was used to evaluate the effects of pressure and tension on cell migration, shape, and α-tubulin architecture in a transformed cell line (DITNC1 rat cortical astrocytes). METHODS: During the assay period, cells were exposed to one of four experimental configurations: (1) control pressure and control gas tension; (2) high-pressure (7.4 mm Hg) and reduced gas tension; (3) control pressure and reduced gas tension; and (4) high-pressure and control gas tension. RESULTS: Calculations suggested that the cells in configurations 2 and 3 were hypoxic, as confirmed by direct measurements in configuration 2. No effects of hydrostatic pressure were observed on cell migration or α-tubulin architecture. However, cells cultured under low gas tension (configurations 2 and 3) showed increased migration at 48 and 72 hours (P < 0.05). CONCLUSIONS: A hydrostatic pressure of 7.4 mm Hg has no effect on DITNC1 astrocytes cultured on rigid coverslips, whereas hypoxia associated with a fluid column creating this pressure does. These results differ from those in a previous report, the results of which may be explained by altered gas tensions in the culture medium. Steps are recommended for control of secondary effects when testing the effect of pressure on cultured cells.
Department of Bioengineering, Imperial College London, London, UK.
Classification:
3.6 Cellular biology (Part of: 3 Laboratory methods)
3.9 Pathophysiology (Part of: 3 Laboratory methods)