PMID: 9660497Jul 11, 1998Paper

Autofluorescent inclusions in long-term postconfluent cultures of retinal pigment epithelium

Investigative Ophthalmology & Visual Science
J M Burke, C M Skumatz

Abstract

To examine the accumulation and morphologic features of fluorescent inclusions in retinal pigment epithelial (RPE) cells in vitro and to determine whether accumulation correlates with parameters of age. Cultured human RPE cells were maintained undisturbed at confluence for intervals as long as 2 years and then were examined for autofluorescent inclusions by fluorescence and electron microscopy, with comparisons to the lipofuscin of freshly isolated RPE. Autofluorescence was also examined in bovine RPE cultures that were aged in vitro by replicative senescence. In some postconfluent cultures,the activity of the lysosomal enzyme cathepsin D was measured, and the effect of cell growth on autofluorescence was examined by reinducing cell proliferation in late-stage cultures. Autofluorescent inclusions accumulated in human and bovine RPE cultures after extended postconfluent periods. Accumulation was not accelerated in cultures from older donors or in cultures that were aged in vitro. The number of granules per RPE cell varied for lipofuscin in situ and inclusions in vitro, although the latter were more heterogeneous in size and shape and in ultrastructural appearance of the granule contents. Lipofuscin and autofluorescent inclusions...Continue Reading

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