gms | German Medical Science

102. Jahrestagung der DOG

Deutsche Ophthalmologische Gesellschaft e. V.

23. bis 26.09.2004, Berlin

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Identification and localization of corneal epithelial stem cells

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  • corresponding author F. E. Kruse - Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen
  • C. Hofmann-Rummelt - Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen
  • K. Blüthner - Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen
  • B. Seitz - Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen
  • C. Cursiefen - Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen
  • U. Schlötzer-Schrehardt - Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen

Evidenzbasierte Medizin - Anspruch und Wirklichkeit. 102. Jahrestagung der Deutschen Ophthalmologischen Gesellschaft. Berlin, 23.-26.09.2004. Düsseldorf, Köln: German Medical Science; 2004. Doc04dogDO.08.04

The electronic version of this article is the complete one and can be found online at: http://www.egms.de/en/meetings/dog2004/04dog071.shtml

Published: September 22, 2004

© 2004 Kruse et al.
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Outline

Text

Objective

A major problem in corneal stem cell biology is the lack of reliable markers for the identification and exact localization of limbal stem cells. For this purpose, we investigated the usefulness of various epithelial stem cell markers by means of combined immunohistochemical stainings of human tissues in situ.

Methods

Frozen and paraffin sections from 20 donor eyes (age 32-92 years) were immunolabeled with commercially available antibodies against p63, Ki-67, ABCG-2, Connexin 43 (Cx43), alpha-enolase, keratin K3, nestin, PAX-6, beta-catenin, and various integrins by applying single and double labeling techniques.

Results

Staining patterns were largely dependent on tissue processing, antibody source, and individual tissues. Nuclear P63 staining was mainly localized to clusters of limbal basal cells, but also to suprabasal limbal and basal cells of the corneal and conjunctival epithelium, and partly colocalized with the proliferation marker Ki-67. Membranous staining for ABCG-2 was strictly confined to groups of basal cells at the limbus. Subpopulations of limbal basal cells were Cx43- and integrin alpha-6 -negative, whereas the suprabasal cells showed some positivity along their cell membranes. PAX-6 and alpha-enolase represented additional markers with predominant staining of limbal basal cells, whereas nestin, beta-catenin, and other integrins revealed no preferential staining pattern. Double labeling experiments confirmed that clusters of p63- and ABCG-2-positive basal cells at the limbus did neither express Cx43 nor integrin alpha-6.

Conclusions

The existing controversies about corneal stem cell markers may have resulted from interindividual variations in staining patterns and from different methodologies applied. The present findings suggest that these cells can be identified by a combination of various markers. A subpopulation of basal epithelial cells at the limbus expressing p63 and ABCG-2, but not Cx43 and integrin alpha-6, comprises about 10% of basal cells and may represent the corneal epithelial stem cells.