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Apoptosis Gene‐Expression Profiling of Ex Vivo Expanded Human Limbal Epithelial Cells
Author(s) -
Prabhakar Savita,
Utheim Tor Paaske,
Raeder Sten,
Utheim Oygunn Aass,
Cai Yiqing,
Roald Borghild,
Haug Kristiane,
Kvalheim Anders,
Messelt Edvard Berger,
Drolsum Liv,
Zhang Hewen,
Nicolaissen Bjorn,
Lyberg Torstein
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.lb12-b
Subject(s) - apoptosis , ex vivo , dna fragmentation , tunel assay , programmed cell death , biology , andrology , microbiology and biotechnology , gene expression , in vivo , gene , medicine , genetics
Ex vivo expanded human limbal epithelial cells (HLEC) used to treat patients with limbal stem cell deficiency can be preserved in a closed eye bank organ culture (OC) system. This study was conducted to investigate cell death due to apoptosis after eye bank storage of cultured HLEC. HLEC were expanded on intact amniotic membranes and stored for one week at 31°C and 23°C in OC medium or at 5°C in Optisol‐GS. Cultures were fixed in formaldehyde and embedded in paraffin. Low labeling indices were observed under different storage conditions as assessed by immunostaining with caspase‐3 (range 0.2%–1.2%) and a DNA fragmentation assay (TUNEL, range 1.0%–2.3%). Cellular results were confirmed at the gene expression level using the Human Apoptosis RT 2 Profiler™ PCR Array from SuperArray Bioscience, which profiles the expression of key apoptosis genes. The results showed that pro‐apoptotic genes were significantly down regulated and anti apoptotic genes were significantly up regulated in all three storage conditions. This study demonstrates that eye bank storage of cultured HLEC is associated with minor cell death due to apoptosis at both the cellular level and the gene expression level. This work was supported in part by the Eastern Norway Regional Health Authority, the Norwegian Association of the Blind and Partially Sighted and the Blindmission IL.