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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۵۷-۵۷
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عنوان انگلیسی Ps-90: How RPE Cells Generate Giant Colonies on PolyHEMA and Maintain Their Characteristic Expression of Tyrosinase Gene?
چکیده انگلیسی مقاله Objective: Retinal pigment epithelial cells (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. PolyHEMA refer to poly 2 hydroxy ethyl metacrylat. Poly 2 hydroxy ethyl metacrylat (PolyHEMA) is a hydrophobic polymer that does not normally support attachment of mammalian cells. It is the basic component of contact lenses In the current study we investigated effect of PolyHEMA as a cell culture substrate on viable RPE cells and expression of Tyrosinase gene Materials and Methods: A stock solution of poly- HEMA (120 mg/ml) was prepared in 95% ethanol and allowed to be dried under laminar hood then was sterilized by UV and washed by PBS. Human globs were obtained earlier than 24 hours after death from the central eye bank of Islamic Republic of Iran. Neonatal human globes were dissected and RPE cells were isolate and cultured in DMEM: F12 (1:1) supplemented with 10% FBS. Cells between passages 2 and 5 were cultured on PolyHEMA and polystyrene substrates (as control) in 24 wells culture palates. DMEM/F12 was supplemented with 10%FBS or 30%AF or DMEM/F12 serum free media were used to nourish cultured cells on polystyrene or PolyHEMA coated vessels. 2 days later MTT assay (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) was applied to assess the number of cells RNA extraction, cDNA synthesis and real time polymerase chain reaction (RT-PCR) were also performed to evaluate RPE cells marker (Tyrosinase) in cultured RPE cells Results: The number of RPE cells on PolyHEMA and polystyrene were the same and Tyrosinase' expression in RPE Cells was cultured on polyHEMA is more than polystyrene. Conclusion: PolyHEMA is a flexible and hydrophobic polymer that cells can survive on it, form giant, pigmented colonies and be re-cultured .Tyrosinase' expression revealed that PolyHEMA could support RPE cultures to establish their population as the main constituents of the giant colonies.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/894
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