Cellular Chemosensitivity Assays: An Overview

Sumantran, Venil N.

doi:10.1007/978-1-61779-080-5_19

Cited by 142 publications

(144 citation statements)

References 59 publications

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“…Cell viability was evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenil tetrazolium bromide assay (Sigma-Aldrich) as described by Sumantran [29]. Briefly, medium from treated and untreated fibroblasts grown in 96-well plates was replaced by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenil tetrazolium bromide solution and incubated for 3 h at 37°C.…”

Section: Cell Viabilitymentioning

confidence: 99%

Effect of Readthrough Treatment in Fibroblasts of Patients Affected by Lysosomal Diseases Caused by Premature Termination Codons

et al. 2015

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Aminoglycoside antibiotics, such as gentamicin, may induce premature termination codon (PTC) readthrough and elude the nonsense-mediated mRNA decay mechanism. Because PTCs are frequently involved in lysosomal diseases, readthrough compounds may be useful as potential therapeutic agents. The aim of our study was to identify patients responsive to gentamicin treatment in order to be used as positive controls to further screen for other PTC readthrough compounds. With this aim, fibroblasts from 11 patients affected by 6 different lysosomal diseases carrying PTCs were treated with gentamicin. Treatment response was evaluated by measuring enzymatic activity, abnormal metabolite accumulation, mRNA expression, protein localization, and cell viability. The potential effect of readthrough was also analyzed by in silico predictions. Results showed that fibroblasts from 5/11 patients exhibited an up to 3-fold increase of enzymatic activity after gentamicin treatment. Accordingly, cell lines tested showed enhanced well-localized protein and/or increased mRNA expression levels and/or reduced metabolite accumulation. Interestingly, these cell lines also showed increased enzymatic activity after PTC124 treatment, which is a PTC readthroughpromoting compound. In conclusion, our results provide a proof-of-concept that PTCs can be effectively suppressed by readthrough drugs, with different efficiencies depending on the genetic context. The screening of new compounds with readthrough activity is a strategy that can be used to develop efficient therapies for diseases caused by PTC mutations.

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Section: Cell Viabilitymentioning

confidence: 99%

Effect of Readthrough Treatment in Fibroblasts of Patients Affected by Lysosomal Diseases Caused by Premature Termination Codons

et al. 2015

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“…A tetrazolium reagent is reduced by mitochondrial dehydrogenases into blue formazan molecules, which can be measured by spectrophotometry. 12 The amount of formazan produced is directly proportional to the total viable cell number. After each experimental period, the medium was replaced by MTT solution (5 mg/mL) diluted 1:10 in PBS (100 µL/well) and incubated for 3 h at 37°C.…”

Section: Cytotoxicity Assaymentioning

confidence: 99%

Effect of time of extraction on the biocompatibility of endodontic sealers with primary human fibroblasts

2012

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The aim of this work was to evaluate the effects of different times of extraction on the cytotoxicity of six representatives of different root canal sealer groups-Real Seal SE, AH Plus, GuttaFlow, Sealapex, Roth 801, and ThermaSeal Plus-with human gingival fibroblasts. The materials were prepared according to manufacturers' specifications, and were incubated in culture medium (DMEM) at 37°C for 1, 7, 14, 21, and 28 days, with daily washing, to simulate periodontal ligament clearance. Human fibroblasts were exposed to the final extracts at 24 hours, and cell viability was determined by MTT assay, with exposure to unconditioned DMEM as a negative control. Statistical analysis comparing cytotoxicities at each exposure time was performed by ANOVA with Scheffé adjustment for multiple comparisons at a 95% confidence level. Results indicated that GuttaFlow was significantly less cytotoxic than all other sealers (p < 0.05) at 1 day of extraction. After 7 days of extraction, cell viability for GuttaFlow was significantly increased as compared with that of all groups except sealer AH Plus. At day 14, cytotoxicity of Sealapex was significantly higher than that of all other sealers (p < 0.05). At days 21 and 28, there were no significant differences in cytotoxicity among sealer groups. All materials presented some level of cytotoxicity to fibroblasts, while GuttaFlow was the least cytotoxic sealer tested. However, the cytotoxicity of all materials seemed to decrease similarly in a time-dependent manner.

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“…A number of chemosensitivity assays were developed over the last decades to predict the responsiveness of tumors to chemotherapy [19][20][21] . In recent years, the ATP-TCA method is a novel approach to test chemosensitivity in solid tumors [22][23][24][25][26] .…”

Section: Discussionmentioning

confidence: 99%

Heterogeneity of chemosensitivity in esophageal cancer using ATP-tumor chemosensitivity assay

Ling

et al. 2012

Acta Pharmacol Sin

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Aim: Current chemotherapy for esophageal cancer is conducted on the basis of empirical information from clinical trials, which fails to take into account the known heterogeneity of chemosensitivity between patients. This study was aimed to demonstrate the degree of heterogeneity of chemosensitivity in esophageal cancers. Methods: A total of 42 esophageal cancer specimens were collected. The heterogeneity of chemosensitivity in esophageal cancer specimens was examined using an ex vivo ATP-tumor chemosensitivity assay (ATP-TCA). Results: Thirty eight specimens produced evaluable results (90.5%). The most active single agent tested was nedaplatin, to which 28.9% of samples were sensitive. Combinations of chemotherapy agents exhibited much higher sensitivity: cisplatin+paclitaxel was sensitive in 16 of 38 (42.1%) of samples, while nedaplatin+paclitaxel was more effective, which was sensitive in 20 of 38 cases (52.6%). Conclusion: There was a marked heterogeneity of chemosensitivity in esophageal cancer. Chemosensitivity testing may provide a practical method for testing new regimens before clinical trials in esophageal cancer patients.

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Cellular Chemosensitivity Assays: An Overview

Cited by 142 publications

References 59 publications

Effect of Readthrough Treatment in Fibroblasts of Patients Affected by Lysosomal Diseases Caused by Premature Termination Codons

Effect of Readthrough Treatment in Fibroblasts of Patients Affected by Lysosomal Diseases Caused by Premature Termination Codons

Effect of time of extraction on the biocompatibility of endodontic sealers with primary human fibroblasts

Heterogeneity of chemosensitivity in esophageal cancer using ATP-tumor chemosensitivity assay

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