Format

Send to

Choose Destination
Invest Ophthalmol Vis Sci. 2014 Aug 14;55(9):5795-805. doi: 10.1167/iovs.14-14664.

Comparative transcriptomic analysis of cultivated limbal epithelium and donor corneal tissue reveals altered wound healing gene expression.

Author information

1
National Institute for Cellular Biotechnology, Dublin City University, Dublin, Ireland.
2
Irish Blood Transfusion Service, Dublin, Ireland.
3
Hermitage Clinic, Dublin, Ireland.
4
The Royal Victoria Eye and Ear Hospital, Dublin, Ireland Royal College of Surgeons in Ireland, Dublin, Ireland.
5
The Royal Victoria Eye and Ear Hospital, Dublin, Ireland.

Abstract

PURPOSE:

The improved surgical outcomes associated with transplantation of cultivated amniotic membrane expanded limbal epithelium (AMLE) compared to traditional donor methods has led to substantial adoption of this technique for treatment of limbal stem cell deficiency.

METHODS:

The mRNA expression profiles of AMLE and CE were assayed using microarrays. Transcripts with a 1.5-fold change in either direction in addition to a Bonferroni adjusted P value < 0.05 were considered to be differentially expressed. Expression changes detected by microarray profiling and important corneal-limbal markers were assessed using quantitative real-time PCR (qRT-PCR) and immunofluorescence staining.

RESULTS:

A total of 487 probe sets (319 upregulated and 168 downregulated) were found to be differentially expressed between AMLE and CE. Enrichment analysis revealed significant overrepresentation of multiple biological processes (e.g., response to wounding, wound healing, and regulation of cell morphogenesis) within the differentially expressed gene list. The expression of a number of genes that were upregulated (ABCG2, S100A9, ITGA5, TIMP2, FGF5, PDGFC, SEMA3A) and downregulated (KLF4, P63α) in AMLE was confirmed using qRT-PCR. Immunofluorescence confirmed that AMLE cultures were P63α, ABCG2, CK3, CK12, and E-cadherin (E-cad) positive.

CONCLUSIONS:

In this study, we have shown that genes associated with wound healing processes are upregulated in AMLE. These gene expression changes may contribute to corneal restoration and the positive outcomes associated with transplantation.

KEYWORDS:

corneal epithelial; cultivated; limbal; stem cell; wound healing

PMID:
25125605
DOI:
10.1167/iovs.14-14664
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Silverchair Information Systems
Loading ...
Support Center