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TISSUE-SPECIFIC STEM CELLS |
aDepartment of Ophthalmology, Chang Gung Memorial Hospital, Keelung, Taiwan;
bGraduate Institute of Clinical Medical Sciences, Chang Gung University, Kwei-shan, Taoyuan, Taiwan;
cDepartment of Physiology and Pharmacology, Chang Gung University, Kwei-shan, Taoyuan, Taiwan;
dGenomic Medicine Research Core Laboratory (GMRCL) of Chang Gung Memorial Hospital, Kwei-shan, Taoyuan, Taiwan;
eDepartment of Biotechnology, Ming-Chuan University, Kwei-shan, Taoyuan, Taiwan
Key Words. Limbal epithelial cells • Amniotic membrane • Interleulin-1 receptor antagonist • Cytokines • Extracellular matrix
Correspondence: Chuen-Mao Yang, Ph.D., Department of Physiology and Pharmacology, Chang Gung University, 259 Wen-Hwa 1st Road, Kwei-San, Tao-Yuan, Taiwan. Telephone: 03-2118800 (ext. 5123); Fax: 03-2118365; e-mail: chuenmao{at}mail.cgu.edu.tw
Received November 25, 2005;
accepted for publication May 22, 2006.
First published online in STEM CELLS EXPRESS June 1, 2006.
Stem cells of the corneal epithelium have been found to be located exclusively at the anatomical junction between the cornea and the conjunctiva, the limbus. Ex vivo expanded limbal epithelial cells on amniotic membrane (AM) are capable of restoring the corneal surface with limbal stem cell deficiency. Recent studies indicate that intact AM preserves the limbal epithelial phenotype and that distinct epithelial morphology is noted among various culture matrix. However, the factors in response to the interaction between limbal epithelial cells and AM were not well understood. Using Annexin V-fluorescein isothiocyanate staining, we found that human limbal epithelial cells expanded on intact human AM demonstrated fewer apoptotic cells as compared with those on plastic dishes. To identify the anti-apoptotic factors, we performed cDNA microarray analysis and showed that interleukin-1 receptor antagonist (IL-1RA) was overexpressed in cultures on intact AM, which was confirmed by reverse transcription-polymerase chain reaction (RT-PCR), real-time quantitative PCR (Q-PCR) and enzyme-linked immunosorbent assay. In addition, we also noted that the phenomenon of apoptosis detected in cultures on plastic dishes could be reversed by adding recombinant IL-1RA protein into the media, whereas apoptosis of limbal epithelial cells cultivated on intact AM could be induced by exogenous neutralizing IL-1RA neutralizing antibody. These results demonstrated that intact human AM may prevent cultured human limbal epithelial cells from undergoing apoptosis. IL-1RA might be a candidate mediator to exert as an anti-apoptotic molecule during the interaction between human limbal epithelial cells and intact human AM.
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