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Kenji Inoue, Shunichiro Kubota, Tadahiko Tsuru, Makoto Araie, Yousuke Seyama; Cholestanol Induces Apoptosis of Corneal Endothelial and Lens Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2000;41(5):991-997.
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purpose. To determine whether cholestanol induces cornea endothelial and lens
epithelial cell death in vitro.
methods. Cornea endothelial and lens epithelial cells were cultured in minimum
essential media with 10% fetal bovine serum containing 10 μg/ml
cholesterol in ethanol, 10 μg/ml cholestanol in ethanol, or 1%
ethanol. These cells, stained using the terminal deoxynucleotidyl
transferase (TdT) dUTP nick-end labeling (TUNEL) method, were analyzed
by laser cytometer. The activities of ICE and CPP32 proteases in cells
were also measured.
results. Both cornea endothelial and lens epithelial cells cultured with 10μ
g/ml cholestanol showed a significant loss of viability. The nuclei
of these cells cultured with 10 μg/ml cholestanol were more
frequently stained than those exposed to 10 μg/ml cholesterol or 1%
ethanol. Quantitative analysis of apoptotic DNA fragmentation confirmed
that the cholestanol induced apoptosis of these cells in a
time-dependent manner. The activities of interleukin-1β–converting
enzyme (ICE) and CPP32 proteases for cells cultured with 10 μg/ml
cholestanol were significantly higher than those observed in control
conclusions. In vitro, cholestanol was taken up by corneal endothelial cells and
lens epithelial cells, an event that led to apoptosis of these
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