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Q. Peng, L. Wang, X. Cheng, P. Ren; Modified Method of Isolating and Culture Rabbit Retinal Pigment Epithelial Cells in vitro . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3043.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To investigate an effective method of isolating and culture rabbit Retinal Pigment Epithelial (RPE) cells in vitro. Methods: RPE cells were isolated from day 1 rabbit retina by two–step enzyme dissociation method. The first–step enzymes (225u/ml hyaluronidase combined 0.125% trypsin) was used to loosen the connection between RPE cells and the neural retina. The second–step enzyme (0.25% trypsin) was used to dissociate RPE cells into single cell. RPE cells isolated were expanded in serum free define 1640 media. Cells at passage 1 were subjected to assay about their quantity, purity, survival rate, growth ability and morphologic characteristics with some methods (RPE sheet assay, H–E stains, Immunohistochemical method, light and electron microscopy). Results: Isolated rabbit RPE cells with modified method have a lot of advantages – intact cell sheet morphology, higher purity, high quantity, strong growth ability (a lot of binuclear cells), short converging date and normal RPE cells morphologic characteristics. Conclusions: This method is efficient and feasible, these cells could be used for some researches about cell biology, cellular pharmacology, genetic expression, cell light– damage and cellular apoptosis etc.
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