November 1985
Volume 26, Issue 11
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Articles  |   November 1985
An improved method for isolation and culture of rat retinal pigment epithelial cells.
Investigative Ophthalmology & Visual Science November 1985, Vol.26, 1599-1609. doi:
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      P L Mayerson, M O Hall, V Clark, T Abrams; An improved method for isolation and culture of rat retinal pigment epithelial cells.. Invest. Ophthalmol. Vis. Sci. 1985;26(11):1599-1609.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Retinal pigment epithelial cells from normal, Long Evans (LE) and retinal dystrophic (RCS) rats can be grown in vitro (Edwards, 1977). An improved technique is described which permits a more rapid isolation of RPE cells, and routinely gives high cell yields (30,000-40,000/eye), excellent cell viability (95%), and high plating efficiencies (95-100%). Whole eyes are treated with hyaluronidase and collagenase followed by trypsin. These enzymes degrade components of the extracellular matrix, releasing sheets of RPE from adherent attachments to the retina and choroid. Trypsin was then used to dissociate the sheets into single cells. RPE cells are grown to confluence in primary culture. This technique permits RPE cell isolation from both normal and retinal dystrophic (RCS) rats, 8-15 days of age. Normal cells isolated by this technique consistently show excellent phagocytosis in vitro.

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