مجله دانشکده پزشکی اصفهان (Nov 2014)
The Effect of Polyhydroxyethylmethacrylate (Poly-HEMA), as a Cell Culture Substrate, on Viable and Preserving Nature of Retinal Pigment Epithelium (RPE) Cells
Abstract
Background: Retinal pigment epithelium (RPE) cells play an important role in the maintenance of normal function of the retina. Tyrosinase is a marker of RPE cells that functions in synthesis of melanin. Polymer of polyhydroxyethylmethacrylate (Poly-HEMA) is the basic component of contact lenses. We investigated the effect of Poly-HEMA, as a cell culture substrate, on viable and preserving nature of RPE cells. Methods: RPE cells between passages 2 and 5 were cultured on Poly-HEMA (12 mg/ml) in 24 wells culture palates. Dulbecco's modified Eagle's medium (DMEM/F12) + 10% Fetal bovine serum (FBS) were used to nourish cultured cells on polystyrene and Poly-HEMA coated vessels. Morphology, the rate of cell proliferation and cell death of RPE cells cultured on Poly-HEMA polymer were evaluated in periodic times. Real-time polymerase chain reaction (RT-PCR) was performed to evaluate tyrosinase in cultured RPE cells. Findings: The cells cultured on Poly-HEMA formed many colonies. These colonies could be re-cultured on polystyrene. The number of RPE cells on Poly-HEMA and polystyrene were the same while the proliferation rate of the cultured cells on Poly-HEMA was reduced; no remarkable cell death was detected on polymer and on polystyrene plates. The giant colonies which were formed on Poly-HEMA were re-cultured. The expression of tyrosinase gene was detected 434.65 in cultured RPE cells on the polymer. Conclusion: Poly-HEMA is a hydrophobic polymer. When RPE cells were cultured on Poly-HEMA they could not adhere to substrate and formed many colonies. Based on our study, Poly-HEMA did not induce cell death in RPE cells but the proliferation of cells was reduced. Tyrosinase expression revealed that Poly-HEMA could support RPE cultures to establish their population as the main constituents of the giant colonies.
