May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Characterization of a spontaneously human Müller cell line generated from a diabetic donor
Author Affiliations & Notes
  • C. Lupien
    Unite Recherche Ophtalmologie, Centre Recherche CHUL, Université Laval, Ste–Foy, PQ, Canada
  • C. Salesse
    Unite Recherche Ophtalmologie, Centre Recherche CHUL, Université Laval, Ste–Foy, PQ, Canada
  • Footnotes
    Commercial Relationships  C. Lupien, None; C. Salesse, None.
  • Footnotes
    Support  Canadian Institutes of Health Research (CIHR)
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 5297. doi:
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      C. Lupien, C. Salesse; Characterization of a spontaneously human Müller cell line generated from a diabetic donor . Invest. Ophthalmol. Vis. Sci. 2004;45(13):5297.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: Müller cells are the predominant type of glial cells in the retina. These cells play very important functions in the retina. Alterations in Müller cell behavior are observed in retinal tissue from patients with various retinal disorders, including proliferative diabetic retinopathy. The purpose of this study was to compare normal human Müller cells with a spontaneously human Müller cell line generated from a diabetic donor. Methods: A spontaneously human Müller cell line (HMCL) was obtained from a 69 years old diabetic donor. These transformed cells are shown to be distinguishable from normal human Müller cells by their morphology as well as on the basis of immunofluorescence analyses. This cell line was investigated for the expression of known markers of Müller cells by immunofluorescence, including cellular retinaldehyde binding protein (CRALBP), glutamine synthetase and glial fibrillary acidic protein (GFAP). Since the morphology of HMCL was not typical of Müller cells, immunofluorescence analyses were also performed with known markers for epithelial cells (cytokeratins K8/K18) and endothelial cells (von Willebrand factor and platelet–endothelial cell adhesion molecule). Results: In contrast to the typical fibroblast–like morphology of Müller cells, HMCL showed an epithelial shape. However, HMCL grow as clusters which is characteristic of proliferative Müller cells. Immunofluorescence analyses showed that both normal human Müller cells and HMCL express CRALBP, glutamine synthetase and GFAP. In addition, HMCL express the cytokeratins K8/K18. However, they do not express the typical markers for endothelial cells. Finally, HMCL have reached a large number of passages without any change in its morphology or expression of markers compared to Müller cells which typically can not be passed beyond a small number of passages. Conclusions: The epitheloid–like morphology of HMCL is consistent with the expression of cytokeratins K8/K18 which are not normally found in Müller cells. However, HMCL are negative for endothelial cell markers and positive for all known markers of Müller cells. These data thus suggest that HMCL correspond to Müller cells although they bear some epitheloid–like characteristics. SAGE studies are in progress to compare gene expression in these cell populations.

Keywords: Muller cells • diabetes • retinal culture 
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