Mouse retinal pigmented epithelial cell lines retain their phenotypic characteristics after transfection with human papilloma virus

A new tool to further the study of RPE biology

Paola Catanuto, Diego Gabriel Espinosa Heidmann, Simone Pereira-Simon, Patricia Sanchez, Pedro Salas, Eleut Hernandez, Scott W. Cousins, Sharon J. Elliot

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Development of immortalized mouse retinal pigmented epithelial cell (RPE) lines that retain many of their in vivo phenotypic characteristics, would aid in studies of ocular diseases including age related macular degeneration (AMD). RPE cells were isolated from 18-month-old (estrogen receptor knockout) ERKOα and ERKOβ mice and their C57Bl/6 wildtype littermates. RPE65 and cellular retinaldehyde binding protein (CRALBP) expression, in vivo markers of RPE cells, were detected by real-time RT-PCR and western analysis. We confirmed the presence of epithelial cell markers, ZO1, cytokeratin 8 and 18 by immunofluorescence staining. In addition, we confirmed the distribution of actin filaments and the expression of ezrin. To develop cell lines, RPE cells were isolated, propagated and immortalized using human papilloma virus (HPV) 16 (E6/E7). RPE-specific markers and morphology were assessed before and after immortalization. In wildtype littermate controls, there was no evidence of any alterations in the parameters that we examined including MMP-2, TIMP-2, collagen type IV, and estrogen receptor (ER)α and ERβ protein expression and ER copy number ratio. Therefore, immortalized mouse RPE cell lines that retain their in vivo phenotype can be isolated from either pharmacologically or genetically manipulated mice, and may be used to study RPE cell biology.

Original languageEnglish (US)
Pages (from-to)99-105
Number of pages7
JournalExperimental eye research
Volume88
Issue number1
DOIs
StatePublished - Jan 5 2009
Externally publishedYes

Fingerprint

Papillomaviridae
Transfection
Cell Biology
Epithelial Cells
Cell Line
Estrogen Receptors
Collagen Receptors
Keratin-8
Retinaldehyde
Keratin-18
Tissue Inhibitor of Metalloproteinase-2
Collagen Type IV
Eye Diseases
Macular Degeneration
Matrix Metalloproteinases
Actin Cytoskeleton
Fluorescent Antibody Technique
Real-Time Polymerase Chain Reaction
Carrier Proteins

Keywords

  • estrogen receptors
  • extracellular matrix
  • immortalization
  • mouse retinal pigmented epithelial cells

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Mouse retinal pigmented epithelial cell lines retain their phenotypic characteristics after transfection with human papilloma virus : A new tool to further the study of RPE biology. / Catanuto, Paola; Espinosa Heidmann, Diego Gabriel; Pereira-Simon, Simone; Sanchez, Patricia; Salas, Pedro; Hernandez, Eleut; Cousins, Scott W.; Elliot, Sharon J.

In: Experimental eye research, Vol. 88, No. 1, 05.01.2009, p. 99-105.

Research output: Contribution to journalArticle

Catanuto, Paola ; Espinosa Heidmann, Diego Gabriel ; Pereira-Simon, Simone ; Sanchez, Patricia ; Salas, Pedro ; Hernandez, Eleut ; Cousins, Scott W. ; Elliot, Sharon J. / Mouse retinal pigmented epithelial cell lines retain their phenotypic characteristics after transfection with human papilloma virus : A new tool to further the study of RPE biology. In: Experimental eye research. 2009 ; Vol. 88, No. 1. pp. 99-105.
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