ARPE-19, a human retinal pigment epithelial cell line with differentiated properties

K. C. Dunn, A. E. Aotaki-Keen, F. R. Putkey, Leonard M Hjelmeland

Research output: Contribution to journalArticle

875 Citations (Scopus)

Abstract

The retinal pigment epithelium (RPE) plays a critical role in the development and maintenance of adjacent photoreceptors in the vertebrate retina. This study describes the development and characterization of ARPE-19, a spontaneously arising human RPE cell line with normal karyology which forms polarized epithelial monolayers on porous filter supports. The cell line was established by selective trypsinization of a primary RPE culture resulting in a uniform population of highly epithelial cells which exhibit a strong growth potential. To determine the extent of biochemical differentiation, the expression of the RPE-specific markers CRALBP and RPE65 was examined by Northern analysis. A single 1.6 kb CRALBP mRNA transcript and a single 2.8 kb RPE65 transcript were detected in samples of ARPE-19 total mRNA. The expression of CRALBP protein in ARPE-19 cell lysate was detected by Western blot analysis and immunocytochemistry was used to detect CRALBP throughout the cytoplasm of most, but not all, cells in confluent cultures. The essential criteria for monolayer formation were determined experimentally and it was found that ARPE-19 cells exhibit morphological polarization when plated on laminin-coated Transwell-COL filters in medium with a low serum content. The time course of tight-junction formation was determined by recording the transepithelial resistance of monolayers and reached a maximum of 50-100 Ω cm2 after 4 weeks of culture. Barrier properties of ARPE-19 monolayers were evaluated by measuring the flux of 3H-inulin from the apical to the basolateral compartment of cell culture chambers. Finally, ARPE-19 clonal sublines were generated by serial dilution in an attempt to produce a subline with a high transepithelial resistance (TER). The morphology of the sublines was variable and the cloned cells exhibited a tendency to senesce in culture, confirming that this cell line is not transformed. No subline monolayers developed a TER greater than those recorded for the parent cells. Our results demonstrate that ARPE-19 has structural and functional properties characteristic of RPE cells in vivo and suggest that this cell line will be valuable for in vitro studies of retinal pigment epithelium physiology.

Original languageEnglish (US)
Pages (from-to)155-169
Number of pages15
JournalExperimental Eye Research
Volume62
Issue number2
DOIs
StatePublished - Feb 1996

Fingerprint

Retinal Pigments
Retinal Pigment Epithelium
Epithelial Cells
Cell Line
Messenger RNA
Vertebrate Photoreceptor Cells
Inulin
Tight Junctions
Laminin
Retina
Cytoplasm
Cell Culture Techniques
Western Blotting
Immunohistochemistry
Maintenance
Growth
Serum
Population
Proteins

Keywords

  • Cell line
  • CRALBP
  • Permeable filter
  • REP65
  • Retinal pigment epithelium (RPE)

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

ARPE-19, a human retinal pigment epithelial cell line with differentiated properties. / Dunn, K. C.; Aotaki-Keen, A. E.; Putkey, F. R.; Hjelmeland, Leonard M.

In: Experimental Eye Research, Vol. 62, No. 2, 02.1996, p. 155-169.

Research output: Contribution to journalArticle

Dunn, K. C. ; Aotaki-Keen, A. E. ; Putkey, F. R. ; Hjelmeland, Leonard M. / ARPE-19, a human retinal pigment epithelial cell line with differentiated properties. In: Experimental Eye Research. 1996 ; Vol. 62, No. 2. pp. 155-169.
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