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PMID: 16049325 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Intrastromal invasion by limbal epithelial cells is mediated by epithelial-mesenchymal transition activated by air exposure.

The American journal of pathology ·Vol. 167 ·No. 2 ·2005-08-00 ·Pages 381-93

Kawakita T, Espana EM, He H, Li W, Liu CY, Tseng SC

Abstract

Corneal epithelial stem cells are located in the basal layer of the limbus between the cornea and the conjunctiva. Regulation of these limbal epithelial progenitor cells by the stromal niche dictates corneal surface health. To further characterize this process, limbal explants were cultured at the air-fluid interface, termed air-lifting, to stimulate the niche. As compared to submerged cultures, air-lifting significantly promoted epithelial stratification, migration, proliferation, and intrastromal invasion by limbal epithelial cells. Epithelial intrastromal invasion was noted when the limbal, but not corneal, epithelium was recombined with the limbal stroma containing live, but not dead, cells. Invading limbal basal cells displayed up-regulated nuclear expression of p63 and Ki67, down-regulated E-cadherin and cornea-specific keratin 3, and switched expression of beta-catenin from intercellular junctions to the nucleus and cytoplasm, indicating the activation of the Wnt/beta-catenin pathway. Invaded cells isolated by collagenase from the stroma of air-lifted, but not submerged, explants showed vivid clonal growth on 3T3 fibroblast feeder layers and complete epithelial-mesenchymal transition by expressing nuclear p63 and cytoplasmic S100A4. These findings collectively suggest that epithelial-mesenchymal transition via the Wnt/beta-catenin pathway influences the fate of limbal epithelial cells, likely to be progenitor cells, between regeneration and fibrosis when the stromal niche is activated.

MeSH Terms
Air Animals Cadherins/metabolism Cell Differentiation Cell Movement/physiology Cell Proliferation Cells, Cultured Corneal Stroma/metabolism,pathology Cytoskeletal Proteins/metabolism Epithelial Cells/metabolism,pathology Epithelium, Corneal/metabolism,pathology Fibroblasts/metabolism,pathology Intercellular Signaling Peptides and Proteins/metabolism Keratins/metabolism Ki-67 Antigen/metabolism Limbus Corneae/metabolism,pathology Mesoderm/cytology Mice Phosphoproteins/metabolism Rabbits Trans-Activators/metabolism Wnt Proteins beta Catenin
Chemicals
CTNNB1 protein, mouse Cadherins Cytoskeletal Proteins Intercellular Signaling Peptides and Proteins Ki-67 Antigen Phosphoproteins Trans-Activators Trp63 protein, mouse Wnt Proteins beta Catenin Keratins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kawakita Tetsuya
TissueTech, Inc., Ocular Surface Center, 7000 SW 97 Ave., Suite 213, Miami, FL 33173, USA.
Espana Edgar M
He Hua
Li Wei
Liu Chia-Yiang
Tseng Scheffer C G
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Article Info
Journal
The American journal of pathology
Abbr.
Am J Pathol
ISSN
0002-9440
DOI
10.1016/S0002-9440(10)62983-5
Published
2005-08-00
Pages
381-93
Language
English
Region
United States
NLM ID
0370502
PMCID
PMC1350963
Subset
IM
Grants
NEI NIH HHS · EY15735 · United States
NEI NIH HHS · R01 EY006819 · United States
NEI NIH HHS · EY12486 · United States
NEI NIH HHS · R01 EY015735 · United States
NEI NIH HHS · EY06819 · United States
NEI NIH HHS · R01 EY012486 · United States
External Links
PubMed source DOI: 10.1016/S0002-9440(10)62983-5 PMC Full Text View journal details
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