May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
A New Method for Quantifying Limbal Stem Cell Markers
Author Affiliations & Notes
  • R. M. Corrales
    CIBER-BBN, Valladolid, Spain
    Ocular Surface Group, IOBA, University of Valladolid, Valladolid, Spain
  • A. De la Mata
    Ocular Surface Group, IOBA, University of Valladolid, Valladolid, Spain
  • M. Lopez
    Ocular Surface Group, IOBA, University of Valladolid, Valladolid, Spain
  • V. Saez
    Ocular Surface Group, IOBA, University of Valladolid, Valladolid, Spain
  • M. De la Paz
    Centro de Oftalmología Barraquer, Barcelona, Spain
  • M. Calonge
    CIBER-BBN, Valladolid, Spain
    Ocular Surface Group, IOBA, University of Valladolid, Valladolid, Spain
  • Footnotes
    Commercial Relationships  R.M. Corrales, None; A. De la Mata, None; M. Lopez, None; V. Saez, None; M. De la Paz, None; M. Calonge, None.
  • Footnotes
    Support  Ministerio de Sanidad y Consumo, Instituto de Salud Carlos III: CIBER-BBN CB06/01/0003 and Red Temática de Investigación Cooperativa en Salud. Red TerCel. Federación de Cajas de Ahorros
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 6074. doi:https://doi.org/
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    • Get Citation

      R. M. Corrales, A. De la Mata, M. Lopez, V. Saez, M. De la Paz, M. Calonge; A New Method for Quantifying Limbal Stem Cell Markers. Invest. Ophthalmol. Vis. Sci. 2008;49(13):6074. doi: https://doi.org/.

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

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Abstract

Purpose: : To analyze the expression of 84 genes related to the identification, growth, maintenance, and differentiation of stem cells using a real time reverse transcription (RT2) PCR array (The Human Stem Cell RT² ProfilerTM, SuperArray Bioscience Co.) in healthy adult human corneal and limbal samples.

Methods: : RNA was isolated from 11 limbal and central cornea epithelial cells. cDNA was synthesized and used for a RT2 PCR array, that combines real time PCR sensitivity and specificity with the ability of microarrays to detect the expression of multiple genes simultaneously. The expression of 84 human genes related to: 1) stem cell specific markers (cell cycle regulators, chromosome and chromatin modulators, genes regulating symmetric/asymmetric cell division, self-renewal markers, cytokines and growth factors, genes regulating cell-cell communication, cell adhesion molecules and metabolic markers); 2) stem cell differentiation markers (embryonic, hematopoietic, mesenchymal and neural cell lineage markers); and 3) signalling pathways important for stem cell maintenance (Notch and Wnt pathways).The comparative Ct method was used for analyzing the results using the cornea as calibrator.

Results: : RT2 PCR array indicated an up-regulation in the expression of most limbal genes compared to corneal genes. The relative fold in limbal samples was higher than 7, 4, and 3 for the 9 embryonic, 6 hematopoietic and 6 mesenchymal cell lineage markers, respectively. The highest folds were found for the following genes: CD8A:12.06, DLL3:12.10, ALPI:14.02, ABCG2:15.23, CCND2:15.43, COL1A1:21.83, IGF-1:31.83, K15:87.88 and CXCL12:140.99.

Conclusions: : We found new putative limbal stem cell marker genes by RT2 PCR array which require further investigation.

Keywords: cornea: epithelium • gene microarray • cornea: basic science 
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