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K. Katsumura, M. Kobayashi, T. Ikeda; Spontaneous Establishment of a Novel Human Cell Line Derived From Lens Epithelium . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1226.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: We report the spontaneous establishment of a human lens epithelial cell line and its biochemical characteristics. Methods: The cells obtained from a surgical specimen of cataract were cultured in vitro. Since those cells were subcultured over three years, it was regarded as an established cell line and named human lens epithelial cell-1 (HLEC-1). We researched morphology, karyotype, genotype and immunohistochemistry of HLEC-1 and analyzed the production of extracellular matrix. HLEC-1 was observed under an inverse-type light microscope and an electron microscope. To confirm that the cell line originated from human, chromosomes, prepared from HLEC-1 using trypsin-Giemsa banding techniques were analyzed and nucleotide sequence of the myoglobin gene was determined using a commercial DNA sequencer. The amount of type I and type IV collagens, alfa-A-crystalline and alfa smooth muscle actin production by HLEC-1 under various concentration of cytokines were measured using CC-EIA technique (Katsumura et al. ARVO2002). Results: Under a light microscope, HLEC-1 was spindle/ dendrite-shaped and morphologically similar to lens epithelial cells in vitro primary culture. Electron microscopic observation of HLEC-1 showed that the cell was mitochondria-rich and less number of rough endoplasmic reticulum compared with fibroblastic cell line, WI-38. The karyotype of HLEC-1 was 45X, and the nucleotide sequence of the myoglobin gene was completely identical to that of humans. The production of alfa-A-crystalline by the cell line was demonstrated by immunohistochemistry. The production of type I and type IV collagens, alfa-A-crystalline and alfa-smooth muscle actin were increased by transforming growth factor beta1 and beta2 and inhibited by tranilast, ketotifen fumarate and sodium diclofenac. On the contrary, betamethasone didn’t affect. Conclusions: HLEC-1 is spontaneously established human lens epithelial cell line without the use of exogenous gene transduction. Therefore this cell line is useful as an in vitro model of lens disease and HLEC-1 could be used in studies of anti-after-cataract agents.
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