This Article  • Full text  • PDF  • Reply to article  • Send to a friend  • Save in My Folder  • Save to citation manager  • Permissions  Citing Articles  • Citation map  • Citing articles on HighWire  • Citing articles on ISI (32)  • Contact me when this article is cited  Related Content  • Similar articles in this journal  Topic Collections  • Ophthalmology, Other  • Alert me on articles by topic

Martin Grueterich, MD; Scheffer C. G. Tseng, MD, PhD

Arch Ophthalmol. 2002;120:783-790.

Background  The transplantation of human limbal epithelium on amniotic membrane as a substrate is a new treatment for limbal stem cell deficiency. Limbal epithelial stem cells are characterized by a slow cell cycle and the lack of K3 keratin and connexin 43 (Cx43), a gap junction protein. We investigated Cx43 expression, gap junction intercellular communication (GJIC), and proliferative activity of limbal epithelium expanded on amniotic membrane.

Methods  Connexin 43 expression and bromodeoxyuridine (BrdU) incorporation were determined by immunohistology. The GJIC was investigated by a scrape-loading dye transfer assay. Expression of Cx43 and K3 keratin as well as BrdU-retaining nuclei were also analyzed after xenotransplantation in nude mice.

Results  Limbal epithelium showed mean ± SD 12.4% ± 14.5% positive units of Cx43 expression and a low BrdU labeling index of 2.4% ± 0.9% (n = 5), of which the latter was due to slow cycling, as proved by its increase to 62.0% ± 9.5% after continuous BrdU labeling for 5 days. Most of the expanded epithelium did not show GJIC (83%), significantly more than that grown on plastic (6%; P<.002). Basal cells of the stratified epithelium after xenotransplantation did not express Cx43 and K3 keratin, but their nuclei retained BrdU.

Conclusion  These results support the hypothesis that intact amniotic membrane preferentially preserves and expands Cx43-negative, keratin K3–negative, and GJIC-deficient limbal epithelium, a phenotype resembling that of stem cell–containing limbal basal epithelial cells in vivo.

Clinical Relevance  Intact amniotic membrane is a suitable substrate for bioengineering limbal epithelia for ocular surface reconstruction.

From the Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami School of Medicine (Dr Grueterich), and Ocular Surface Center and Ocular Surface Research & Education Foundation (Dr Tseng), Miami, Fla.

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES

Effects of Organ Culture and Optisol-GS Storage on Structural Integrity, Phenotypes, and Apoptosis in Cultured Corneal Epithelium
Raeder et al.
IOVS 2007;48:5484-5493.
ABSTRACT | FULL TEXT  

The Fate of Limbal Epithelial Progenitor Cells during Explant Culture on Intact Amniotic Membrane
Li et al.
IOVS 2007;48:605-613.
ABSTRACT | FULL TEXT  

Interleukin-1 Receptor Antagonist (IL-1RA) Prevents Apoptosis in Ex Vivo Expansion of Human Limbal Epithelial Cells Cultivated on Human Amniotic Membrane
Sun et al.
Stem Cells 2006;24:2130-2139.
ABSTRACT | FULL TEXT  

Basement membrane dissolution and reassembly by limbal corneal epithelial cells expanded on amniotic membrane.
Li et al.
IOVS 2006;47:2381-2389.
ABSTRACT | FULL TEXT  

Signaling-Transduction Pathways Required for Ex Vivo Expansion of Human Limbal Explants on Intact Amniotic Membrane
He et al.
IOVS 2006;47:151-157.
ABSTRACT | FULL TEXT  

Epithelial proliferative potential of organ cultured corneoscleral rims; implications for allo-limbal transplantation and eye banking
Shanmuganathan et al.
Br. J. Ophthalmol. 2006;90:55-58.
ABSTRACT | FULL TEXT  

Soluble Lumican Glycoprotein Purified from Human Amniotic Membrane Promotes Corneal Epithelial Wound Healing
Yeh et al.
IOVS 2005;46:479-486.
ABSTRACT | FULL TEXT  

In Vitro Antiangiogenic Activity in Ex Vivo Expanded Human Limbocorneal Epithelial Cells Cultivated on Human Amniotic Membrane
Ma et al.
IOVS 2004;45:2586-2595.
ABSTRACT | FULL TEXT  

Suppression of Corneal Neovascularization by PEDF Release from Human Amniotic Membranes
Shao et al.
IOVS 2004;45:1758-1762.
ABSTRACT | FULL TEXT  

Development of a Conjunctival Epithelial Equivalent with Improved Proliferative Properties Using a Multistep Serum-Free Culture System
Ang et al.
IOVS 2004;45:1789-1795.
ABSTRACT | FULL TEXT  

Epithelial cell characteristics of cultured human limbal explants
Joseph et al.
Br. J. Ophthalmol. 2004;88:393-398.
ABSTRACT | FULL TEXT  

Modulation of Keratin and Connexin Expression in Limbal Epithelium Expanded on Denuded Amniotic Membrane with and without a 3T3 Fibroblast Feeder Layer
Grueterich et al.
IOVS 2003;44:4230-4236.
ABSTRACT | FULL TEXT