Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Ca++-switch induction of RPE differentiation.

Daniel J Rak1, Katherine M Hardy, Glenn J Jaffe

  • 1Department of Ophthalmology, The University of Arizona, Tucson, AZ, USA.

Experimental Eye Research
|November 18, 2005
PubMed
Summary

A novel Ca++-switch protocol revitalizes cultured retinal pigment epithelial (RPE) cells, restoring their differentiated phenotype and improving their utility as a model for studying visual diseases.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Ring Melanoma: from Vitrectomy to Enucleation.

Retinal cases & brief reports·2026
Same author

Geographic Atrophy on Fundus Autofluorescence and Color Fundus Photographs: Association with Deep Visual Sensitivity Losses.

Ophthalmology science·2026
Same author

LONG-TERM EFFICACY AND SAFETY OF PHOTOBIOMODULATION IN DRY AGE-RELATED MACULAR DEGENERATION (LIGHTSITE III: 24-MONTH ANALYSIS).

Retina (Philadelphia, Pa.)·2026
Same author

Levodopa in retinal disease: Dopamine pathways, neuroprotective mechanisms, and clinical evidence.

Survey of ophthalmology·2026
Same author

Standardizing Measurement of Complete Retinal Pigment Epithelial and Outer Retinal Atrophy: Classification of Atrophy Meeting Report 8.

Ophthalmology. Retina·2026
Same author

Standardization of Imaging Criteria for Detecting Macular Fibrosis in Neovascular Age-Related Macular Degeneration.

Ophthalmology science·2026

Area of Science:

  • Cell Biology
  • Ophthalmology
  • Tissue Engineering

Background:

  • Cultured retinal pigment epithelial (RPE) cells are vital models for visual disease research.
  • These cells often lose their in vivo differentiated characteristics during culture, limiting their scientific application.

Purpose of the Study:

  • To investigate a calcium switch (Ca++-switch) protocol for restoring the differentiated phenotype of cultured RPE cells.
  • To assess the impact of this protocol on RPE-specific markers and morphology.

Main Methods:

  • Primary RPE cells from adult donors were cultured and subjected to a Ca++-switch protocol.
  • Cells were maintained at confluence for up to 4 months, with paired controls.
  • Phenotype, pigmentation, and expression of tyrosinase, CRABP, myocilin, and bestrophin were analyzed via western blot.

Related Experiment Videos

Main Results:

  • The Ca++-switch protocol induced N-cadherin localization to lateral borders and tyrosinase expression by day 4.
  • Visible pigment accumulation occurred after 8 weeks, and CRABP expression was noted at 12 weeks.
  • Myocilin was detected at 4 months, but bestrophin remained undetectable.

Conclusions:

  • The Ca++-switch protocol effectively promotes RPE cell differentiation and epithelial morphogenesis.
  • Differences in cell spreading and cadherin adhesion were observed, suggesting multiple cell signaling pathways are involved.
  • This method offers a promising approach to enhance the in vitro modeling of RPE biology and disease.