PURPOSE. To investigate how the cellular behavior of cultured retinal pigment epithelial (RPE) cells was affected by the manipulation of early focal contact. METHODS. To manipulate early focal contact, a reduced focal cell-substrate contact area on the micropatterned surfaces was implemented by microfabrication with polydimethylsiloxane (PDMS). The micropatterned PDMS surfaces had a circular pillar with a diameter of 5 μm. The human retinal pigment epithelial cell line, ARPE-19, was seeded onto the fibronectin-coated PDMS surfaces. Cell adhesion, growth, cell cycle, morphology, and interleukin-6 (IL-6) expression were observed, for 3 weeks. RESULTS. The fluorescent images of localized vinculin. and actin stress fibers appeared to be more prominent on smooth PDMS surfaces. Although there was no significant effect on cell adhesion, a statistically significant inhibition of cell cycle progression was observed for micropatterned PDMS surfaces. Similarly, micropatterned surfaces showed significantly less cell growth than that of smooth surfaces. Cultures over a period of 3 weeks showed a distinct cell-cell phenotype discrepancy. Furthermore, IL-6 mRNA and secreted protein induced by IL-1β in ARPE-19 were downregulated on micropatterned PDMS surfaces. CONCLUSIONS. Disturbed focal contact in ARPE-19 cells grown on micropatterned surfaces altered cell cycle, growth, morphology, and the expression of IL-6 in vitro.
ASJC Scopus subject areas
- Sensory Systems
- Cellular and Molecular Neuroscience