Migration and Proliferation Inhibition of ARPE Cells under Ascorbic Acid
Heckelen A.1, Wöltje M.1, Kondring B.2, Jahnen-Dechent W.1, Schrage N. F.2
1IZKF Biomat, Aachen; 2Augenklinik, Universitätsklinikum Aachen
Purpose: Proliferation and migration are essential parameters of proliferative vitreoretinopathy (PVR). Application of cytostatic drugs are related to side effects but since the daunomycine study established in surgical PVR treatment Ascorbic acid has given evidence to act as natural proliferation inhibitor. Another factor of PVR is the migration of cells into the vitreous body. We looked if this parameter is influenced by ascorbic acid levels.
Method: For proliferation determination ARPE cells were incubated in vitro with increasing concentrations of ascorbic acid (0.5-4 mMol, pH 7,4). Cell proliferation was measured by BrdU assay. After replacement of the ascorbic acid containing tissue culture medium (TCM) by normal TCM we measured re-proliferation after 24 hours. With 1mMol ascorbic acid stimulated cells (150000Z/ml) and unstimulated cells for negative control were seeded for migration assay on cell culture inserts consisting of polycarbonat membrane (poresize=8µm). The inserts were transfered into wells of a 24 well plate and incubated for 3h allowing cells to attach on the dry surface of culture insert. Both MCP-1 (monocyte chemotactic protein-1) containing medium (20ng/ml) and pure medium for negative control were injected under the membrane after incubation and cells were
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