Use of the ARPE-19 cell line as a model of RPE polarity: basolateral secretion of FGF5

Invest Ophthalmol Vis Sci. 1998 Dec;39(13):2744-9.


Purpose: To determine the polarity of fibroblast growth factor 5 (FGF5) secretions from retinal pigment epithelium (RPE) cells and to examine the viability and utility of the ARPE-19 cell line as a model for the study of RPE polarity.

Methods: Influenza infection and adenovirus-mediated gene transfer were used to deliver and express genes encoding influenza hemagglutinin (HA), p75-NTR (a neurotrophin receptor), low-density lipoprotein (LDL) receptor (LDLR), and FGF5 in confluent monolayers of ARPE-19 cells. The localization of HA, p75-NTR, and LDLR was determined by confocal microscopy. Domain selective biotinylation assays were used to quantitatively determine the polarities of p75-NTR and LDLR. The secretion of FGF5 into the apical and basal media of ARPE-19 cultures was examined by immunoblot analysis of conditioned media.

Results: Hemagglutinin and p75-NTR were found to be localized on the apical surface of infected and transduced ARPE-19 cells. In contrast, LDLR was associated preferentially with the basolateral membrane of ARPE-19 cells. Biotinylation studies indicated that 84% of p75-NTR was present on the apical surface, and 79% of LDLR was basolaterally polarized. Over the course of 6 hours, more than 90% of the total secreted FGF5 protein accumulated in the basolateral media.

Conclusions: ARPE-19 cells exhibit a polarized distribution of cell surface markers when examined by either confocal microscopy or surface-labeling assays. This indicates that the ARPE-19 cell line is a valid model for studies of RPE cell polarity. FGF5, a secreted protein normally produced by RPE cells, is accumulated preferentially in the basal media after only 6 hours, suggesting that it is vectorially secreted from the basolateral surface of ARPE-19 cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Basement Membrane / metabolism
  • Blotting, Western
  • Cell Line
  • Cell Polarity*
  • Cells, Cultured
  • Electrophoresis, Polyacrylamide Gel
  • Fibroblast Growth Factor 5
  • Fibroblast Growth Factors / genetics
  • Fibroblast Growth Factors / metabolism*
  • Hemagglutinin Glycoproteins, Influenza Virus / genetics
  • Hemagglutinin Glycoproteins, Influenza Virus / metabolism
  • Humans
  • Microscopy, Confocal
  • Pigment Epithelium of Eye / metabolism*
  • Pigment Epithelium of Eye / virology
  • Receptor, Nerve Growth Factor
  • Receptors, LDL / genetics
  • Receptors, LDL / metabolism
  • Receptors, Nerve Growth Factor / genetics
  • Receptors, Nerve Growth Factor / metabolism
  • Transfection


  • FGF5 protein, human
  • Hemagglutinin Glycoproteins, Influenza Virus
  • Receptor, Nerve Growth Factor
  • Receptors, LDL
  • Receptors, Nerve Growth Factor
  • Fibroblast Growth Factor 5
  • Fibroblast Growth Factors