Fluorescence properties of autofluorescent granules generated by cultured human RPE cells

Julie Wassell, Steven Ellis, Janice Burke, Mike Boulton

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

PURPOSE. To compare the fluorescence properties of autofluorescent granules generated by retinal pigment epithelial (RPE) cells in vitro with those of the lipofuscin of RPE in vivo. METHODS. Cultured human RPE cells were maintained in basal medium for as long as 1 year, fed rod outer segments (ROS) daily for as long as 56 days, fed ROS in the presence and absence of leupeptin, or fed liposomes consisting of the major phospholipids in ROS. At different time points, cells were examined for overall fluorescence, and their fluorescence spectra were determined. In addition, chloroform-methanol extracts were examined by thin-layer chromatography and compared with those generated from RPE lipofuscin. RESULTS. Autofluorescent granules accumulated in cultured RPE cells, regardless of the presence of an exogenous substrate or the nature of the substrate. The rate of accumulation of autofluorescent granules was greatest in cells fed ROS. The autofluorescent material generated in cultured RPE cells had some spectral similarities with RPE lipofuscin but differed in solubility and chromatographic mobility of their constituent fluorophores. CONCLUSIONS. The autofluorescent granules generated by cultured RPE, even with different specific substrates, differ from lipofuscin granules in vivo, suggesting that additional properties of RPE cells or of the materials they phagocytose are required to produce autofluorescent materials with the characteristics of lipofuscin.

Original languageEnglish (US)
Pages (from-to)1487-1492
Number of pages6
JournalInvestigative Ophthalmology and Visual Science
Volume39
Issue number8
StatePublished - Jul 1998
Externally publishedYes

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Retinal Pigments
Fluorescence
Epithelial Cells
Lipofuscin
Rod Cell Outer Segment
Thin Layer Chromatography
Chloroform
Phagocytosis
Liposomes
Solubility
Methanol
Phospholipids

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Fluorescence properties of autofluorescent granules generated by cultured human RPE cells. / Wassell, Julie; Ellis, Steven; Burke, Janice; Boulton, Mike.

In: Investigative Ophthalmology and Visual Science, Vol. 39, No. 8, 07.1998, p. 1487-1492.

Research output: Contribution to journalArticle

Wassell, Julie ; Ellis, Steven ; Burke, Janice ; Boulton, Mike. / Fluorescence properties of autofluorescent granules generated by cultured human RPE cells. In: Investigative Ophthalmology and Visual Science. 1998 ; Vol. 39, No. 8. pp. 1487-1492.
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N2 - PURPOSE. To compare the fluorescence properties of autofluorescent granules generated by retinal pigment epithelial (RPE) cells in vitro with those of the lipofuscin of RPE in vivo. METHODS. Cultured human RPE cells were maintained in basal medium for as long as 1 year, fed rod outer segments (ROS) daily for as long as 56 days, fed ROS in the presence and absence of leupeptin, or fed liposomes consisting of the major phospholipids in ROS. At different time points, cells were examined for overall fluorescence, and their fluorescence spectra were determined. In addition, chloroform-methanol extracts were examined by thin-layer chromatography and compared with those generated from RPE lipofuscin. RESULTS. Autofluorescent granules accumulated in cultured RPE cells, regardless of the presence of an exogenous substrate or the nature of the substrate. The rate of accumulation of autofluorescent granules was greatest in cells fed ROS. The autofluorescent material generated in cultured RPE cells had some spectral similarities with RPE lipofuscin but differed in solubility and chromatographic mobility of their constituent fluorophores. CONCLUSIONS. The autofluorescent granules generated by cultured RPE, even with different specific substrates, differ from lipofuscin granules in vivo, suggesting that additional properties of RPE cells or of the materials they phagocytose are required to produce autofluorescent materials with the characteristics of lipofuscin.

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