Using MTT Viability Assay to Test the Cytotoxicity of Antibiotics and Steroid to Cultured Porcine Corneal Endothelial Cells

Wang, Hwei-Zu; Chang, Cheng-Hsien; Lin, Chang‐Ping; Tsai, Mi Ching

doi:10.1089/jop.1996.12.35

Cited by 74 publications

(43 citation statements)

References 4 publications

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“…To confirm this, T-HCECs were treated or untreated with an inhibitor of NF-ÎB translocation. Cells treated with sulfasalazine, a potent inhibitor of NF-ÎB translocation that prevents IÎB degradation [6][7][8][9], or SN-50, a specific permeating inhibitor of NF-ÎB nuclear translocation that contains the nuclear localization sequence for the p50 NF-ÎB subunit to promote cell permeability [10], were exposed to UV. It is interesting to note that cellular death was inhibited in T-HCECs treated with sulfasalazine and SN-50.…”

Section: Discussionmentioning

confidence: 99%

“…After UV irradiation of cultured SV40-transfected human corneal epithelial cells (T-HCECs) [5], translocation of NF-ÎB was evaluated with immunocytochemical staining and electrophoretic mobility shift assay (EMSA). We also examined whether sulfasalazine [6][7][8][9] and SN-50 [10], both potent translocation inhibitors of NF-ÎB, had protective effects on UV-induced cellular damage. The goals of this study were to evaluate whether NF-ÎB was related to cellular damage after UV irradiation and to seek the possibility of protecting irradiated cells from complications of UV exposure.…”

Section: Introductionmentioning

confidence: 99%

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Translocation of Nuclear Factor-κB on Corneal Epithelial Cells Induced by Ultraviolet B Irradiation

Lee

Kim²,

Joo³

2005

Ophthalmic Res

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Purpose: This study was performed to elucidate the role of nuclear factor-ĸB (NF-ĸB) in the death of corneal epithelial cells after ultraviolet (UV) irradiation. Methods: Simian virus 40-transfected human corneal epithelial cells (T-HCECs) were used in this study. Cell cultures were irradiated with a UVB (312 nm) source located 10 cm from the bottom of the slides for 10, 20, 30, or 40 s. Cytotoxicity was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Translocation of NF-ĸB was examined by immunocytochemistry using anti-NF-ĸB p65 antibody and electrophoretic mobility shift assay (EMSA). Sulfasalazine and SN-50, specific NF-ĸB inhibitors, were used to confirm the role of NF-ĸB by pretreating samples for 30 min before UV irradiation, after which cytotoxicity and NF-ĸB translocation were evaluated. Results: When T-HCECs were irradiated with UVB, translocation of NF-ĸB was observed with immunocytochemistry. These translocations peaked 2 h after UV irradiation during EMSA. When pretreated with sulfasalazine or SN-50, the translocation of NF-ĸB was blocked. Cellular death after UV irradiation was also markedly blocked by sulfasalazine. Conclusion: These findings suggest that NF-ĸB plays an important role in cellular death after UV irradiation.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Translocation of Nuclear Factor-κB on Corneal Epithelial Cells Induced by Ultraviolet B Irradiation

Lee

Kim²,

Joo³

2005

Ophthalmic Res

View full text Add to dashboard Cite

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“…So it is necessary to study the cytotoxic effects of these fungicides on human skin and liver, therefore the present study was undertaken to study the effect of carbendazim and copper oxychloride on HaCaT and HepG2 cell lines by using the standard MTT cytotoxicity assay. MTT is a tetrazolium salt which is reduced to purple formazan crystals by succinate dehydrogenase [16].…”

Section: Mtt Cell Viability Assaymentioning

confidence: 99%

In-vitro Assessment of Carbendazim and Copper oxychloride cytotoxicity on HaCaT and HepG2 human cell lines

2017

J App Biol Biotech

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The objective of present study was to evaluate the cytotoxic effect of two fungicides i.e. carbendazim and copper oxychloride on human cell lines HaCaT [keratinocyte] and HepG2 [hepatoma cell line]. The cytotoxic study was done by following the standard MTT [3-[4,5-dimethylthiazol-2-yl]-2,5-diphehyltetrazolium bromide] assay along with standard drug. The MTT assay results showed that these two fungicides showed concentration dependent cytotoxicity, upon incubation for 24 hours with concentration ranging 20-450 µg/ml and IC50 values were determined. The cell viability of HaCaT cells decreased from 100% to 35.86% with increase in carbendazim concentration 20-450 µg/ml. Whereas HepG2 cells were sensitive towards Carbendazim treatment with the decrease in viability up to 30.98% at 450 μg/ml. Copper oxychloride was more lethal, causing total cell death of HaCaT cells at 350 μg/ml, and HepG2 at 450 μg/ml and the present work correlate this toxicity caused by these two fungicides on human skin and liver. The study u nveils the cytotoxic effects of these fungicides on the skin as well as on the liver indicating the long term exposure to these compounds may lead to deleterious effects. Further study is needed on to identify the mechanism and pathway involved in the cytotoxic activity of these fungicides on HaCaT and HepG2 cell lines.

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“…Cell viability assay and the viable cells integration As primary hepatocytes in vitro were unable to proliferate, the MTT method was used to evaluate the relative viability of gel entrapped hepatocytes cultured in hollow fibers as previously described (Wang et al 1996). Briefly, at the culture time of 0,24,48,72,96,120,144 h, gel entrapped hepatocytes were respectively extruded from the hollow fibers with a 5 mL-syringe and immerged in 0.65 mL of the MTT-phosphate buffer solution (1.15 mg/mL) in 24-well plates followed by incubation at 37°C for 3 h which was interrupted by a short shaking every 1 h. Then, the MTT-phosphate buffer solution was discarded and 1.5 mL of isopropanol was added to the cells.…”

Section: Measurement Of Gel Contractionmentioning

confidence: 99%

Interaction between hepatocytes and collagen gel in hollow fibers

2009

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Gel entrapment culture of primary mammalian cells within collagen gel is one important configuration for construction of bioartificial organ as well as in vitro model for predicting drug situation in vivo. Gel contraction in entrapment culture, resulting from cell-mediated reorganization of the extracellular matrix, was commonly used to estimate cell viability. However, the exact influence of gel contraction on cell activities has rarely been addressed. This paper investigated the gel contraction under varying culture conditions and its effect on the activities of rat hepatocyte entrapped in collagen gel within hollow fibers. The hepatocyte activities were reflected by cell viability together with liver-specific functions on urea secretion and cytochrome P450 2E1. Unexpectedly, no gel contraction occurred during gel entrapment culture of hepatocyte under a high collagen concentration, but hepatocytes still maintained cell viability and liver-specific functions at a similar level to the other cultures with normal gel contraction. It seems that cell activities are unassociated with gel contraction.Alternatively, the mass transfer resistance induced by the combined effect of collagen concentration, gel contraction and cell density could be a side effect to reduce cell activities. The findings with gel entrapment culture of hepatocytes would be also informative for the other cell culture targeting pathological studies and tissue engineering.

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Using MTT Viability Assay to Test the Cytotoxicity of Antibiotics and Steroid to Cultured Porcine Corneal Endothelial Cells

Cited by 74 publications

References 4 publications

Translocation of Nuclear Factor-κB on Corneal Epithelial Cells Induced by Ultraviolet B Irradiation

Translocation of Nuclear Factor-κB on Corneal Epithelial Cells Induced by Ultraviolet B Irradiation

In-vitro Assessment of Carbendazim and Copper oxychloride cytotoxicity on HaCaT and HepG2 human cell lines

Interaction between hepatocytes and collagen gel in hollow fibers

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