Genomic Aberrations and Cellular Heterogeneity in SV40-Immortalized Human Corneal Epithelial Cells

Yamasaki, Kenta; Kawasaki, Satoshi; Young, Robert D.; Fukuoka, Hideki; Tanioka, Hidetoshi; Nakatsukasa, Mina; Quantock, Andrew J.; Kinoshita, Shigeru

doi:10.1167/iovs.08-2239

Cited by 44 publications

(33 citation statements)

References 45 publications

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“…The decrease in the number of epithelial cell layers and HCK growth rate may be attributed to an altered genomic content after extensive passages, as reported for other SV40-immortalized cell lines (Ray et al, 1990;Stewart and Bacchetti, 1991). In addition, the used HCE cell line is composed of numerous heterogeneous cell populations, since such genomic aberrations randomly occur and may vary among individual cells (Yamasaki et al, 2009).…”

Section: Tablementioning

confidence: 58%

Development of an in vitro ocular test system for the prediction of all three GHS categories

Bartok

Gabel

Zorn-Kruppa

et al. 2015

Toxicology in Vitro

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Section: Tablementioning

confidence: 58%

Development of an in vitro ocular test system for the prediction of all three GHS categories

Bartok

Gabel

Zorn-Kruppa

et al. 2015

Toxicology in Vitro

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“…2280) was obtained from the RIKEN BioResource Center (Tsukuba, Japan). The cells were maintained in the following culture medium: 1 : 1 mixture of Dulbecco's modified eagle medium and nutrient mixture F‐12 supplemented with 5% heat‐inactivated fetal bovine serum, 5 µg ml –1 recombinant human insulin, 10 ng ml –1 recombinant human epidermal growth factor, 0.5% dimethyl sulfoxide 100 units ml –1 penicillin and 100 µg ml –1 streptomycin (Araki‐Sasaki et al ., 1995; Yamasaki et al ., 2009). Cells were grown at 37 °C in a humidified atmosphere of 5% CO 2 in air.…”

Section: Methodsmentioning

confidence: 99%

Predictive performance of the Vitrigel‐eye irritancy test method using 118 chemicals

Yamaguchi

Kojima²,

Takezawa

2015

J of Applied Toxicology

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We recently developed a novel Vitrigel‐eye irritancy test (EIT) method. The Vitrigel‐EIT method is composed of two parts, i.e., the construction of a human corneal epithelium (HCE) model in a collagen vitrigel membrane chamber and the prediction of eye irritancy by analyzing the time‐dependent profile of transepithelial electrical resistance values for 3 min after exposing a chemical to the HCE model. In this study, we estimated the predictive performance of Vitrigel‐EIT method by testing a total of 118 chemicals. The category determined by the Vitrigel‐EIT method in comparison to the globally harmonized system classification revealed that the sensitivity, specificity and accuracy were 90.1%, 65.9% and 80.5%, respectively. Here, five of seven false‐negative chemicals were acidic chemicals inducing the irregular rising of transepithelial electrical resistance values. In case of eliminating the test chemical solutions showing pH 5 or lower, the sensitivity, specificity and accuracy were improved to 96.8%, 67.4% and 84.4%, respectively. Meanwhile, nine of 16 false‐positive chemicals were classified irritant by the US Environmental Protection Agency. In addition, the disappearance of ZO‐1, a tight junction‐associated protein and MUC1, a cell membrane‐spanning mucin was immunohistologically confirmed in the HCE models after exposing not only eye irritant chemicals but also false‐positive chemicals, suggesting that such false‐positive chemicals have an eye irritant potential. These data demonstrated that the Vitrigel‐EIT method could provide excellent predictive performance to judge the widespread eye irritancy, including very mild irritant chemicals. We hope that the Vitrigel‐EIT method contributes to the development of safe commodity chemicals. Copyright © 2015 The Authors. Journal of Applied Toxicology published by John Wiley & Sons Ltd.

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“…This is presumably associated with the random genetic or epigenetic alterations in cells entering the "crisis" stage discussed above. Due to SV40 TAg-mediated interference with the p53 and pRb pathways, these altered cell clones were not effectively eliminated from the cultures [Yamasaki et al, 2009]. Consequently, a masking effect may have occurred, whereby less-differentiated and transformed cell clones with higher proliferation rates overgrew the cultures .…”

Section: Characteristics Of Late-passage Poco83-3 Culturesmentioning

confidence: 99%

Establishment and Characterization of an SV40 Large T Antigen-Transduced Porcine Colonic Epithelial Cell Line

Kaiser

Böttner²,

Wedel³

et al. 2017

Cells Tissues Organs

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Continuous cell lines have become indispensable tools that have enabled investigations into cellular mechanisms by increasing experimental reproducibility and sample availability, and decreasing the use of experimental animals. To facilitate studies of epithelial barrier function of the porcine colon, we aimed to establish an epithelial cell line with an extended replicative capacity. Cells were isolated from the proximal colon of a 3-week-old piglet and transduced using a recombinant retroviral vector construct containing the simian virus 40 large T antigen (SV40 TAg). We established a clonal epithelial cell line, referred to as PoCo83-3, that stably expressed the SV40 TAg, verified at mRNA and protein levels. PoCo83-3 showed epithelial cell-specific features, such as cobblestone-like morphology, dome structure formation, the presence of apical microvilli, and the expression of keratin 18, E-cadherin and the tight junction-associated proteins zonula occludens-1, occludin, and claudin-1. To validate PoCo83-3 as an in vitro model in epithelial barrier research, proinflammatory cytokine-inducible alterations in barrier integrity were demonstrated by incubating the cells with TNF-α and IFN-γ for 48 h. These cytokine treatments promoted a decreased transepithelial electrical resistance. In summary, PoCo83-3 exhibited an extended life span and a differentiated phenotype while maintaining epithelial characteristics. Based on these results, we present this cell line as a valuable in vitro model for investigations of epithelial barrier function in the porcine colon.

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Genomic Aberrations and Cellular Heterogeneity in SV40-Immortalized Human Corneal Epithelial Cells

Abstract: The results indicate that HCE-T cells have an altered genomic content and that they are composed of heterogeneous cell populations. This should be considered when conducting experiments or interpreting the results of studies that use this cell line.

Cited by 44 publications

References 45 publications

Development of an in vitro ocular test system for the prediction of all three GHS categories

Development of an in vitro ocular test system for the prediction of all three GHS categories

Predictive performance of the Vitrigel‐eye irritancy test method using 118 chemicals

Establishment and Characterization of an SV40 Large T Antigen-Transduced Porcine Colonic Epithelial Cell Line

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