Assessment of the predictive capacity of the optimized in vitro comet assay using HepG2 cells

Hong, Youngtae; Jeon, Hye Lyun; Ko, Kyung Yuk; Kim, Joohwan; Yi, Jung-Sun; Ahn, Il Young; Kim, Tae Sung; Lee, Jong Kwon

doi:10.1016/j.mrgentox.2018.01.010

Cited by 14 publications

(7 citation statements)

References 40 publications

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“…The detailed experimental procedure consists of the four steps listed above. The estimated cells are mixed with up to 1% low melting point agarose (LMPA) [79][80][81][82][83][84][85][86], which (in contrast to commonly used agarose) is liquid under normal conditions. The mixture of cells and LMPA is placed on a microscope slide covered with normal melting point agarose (NMPA) in concentration ranging between 0.5% and 1% [79-82, 84, 86, 87].…”

Section: Comet Assaymentioning

confidence: 99%

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An overview of apoptosis assays detecting DNA fragmentation

2018

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Apoptosis has been recognized as a type of programmed cell death connected with characteristic morphological and biochemical changes in cells. This programmed cell death plays an important role in the genesis of a number of physiological and pathological processes. Thus, it can be very important to detect the signs of apoptosis in a study of cellular metabolism. The present paper provides an overview of methods often being used for detecting DNA fragmentation as one of the most specific findings in apoptosis. To date, three routine assays have been developed for detecting DNA fragmentation: DNA ladder assay, TUNEL assay, and comet assay. All these methods differ in their principles for detecting DNA fragmentation. DNA ladder assay detects the characteristic "DNA ladder" pattern formed during internucleosomal cleavage of DNA. Terminal deoxynUcleotidyl transferase Nick-End Labeling (TUNEL) assay detects DNA strand breaks using terminal deoxynucleotidyl transferase catalyzing attachment of modified deoxynucleotides on the DNA strand breaks. Comet assay can be used for detecting nucleus breakdown producing single/double-strand DNA breaks. The aim of this review is to describe the present knowledge on these three methods, including optimized approaches, techniques, and limitations.

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Section: Comet Assaymentioning

confidence: 99%

“…Next, agarose gel electrophoresis is carried out in direct electrical current under low voltage. For DNA staining, ethidium bromide, propidium iodide, DAPI, acridine orange [72], or SYBR staining [77,[83][84][85] can be used. It has been proven that the comets also can be stained with a permanent silver stain [89].…”

Section: Comet Assaymentioning

confidence: 99%

An overview of apoptosis assays detecting DNA fragmentation

2018

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“…From these results, we judged that the HepG2 cell line was more suitable for the Comet assay than the CHL/IU cell line under these study conditions. A standardized comet test using the TK-6 cell line is being performed to confirm genotoxicity in a 2D cell culture system [26], but the HepG2 cell line is also being studied for genotoxicity in a 2D cell culture system [25,41,42]. In addition, we studied as a representative cell line in a 3D culture system to replace animal testing [31][32][33][34].…”

Section: Discussionmentioning

confidence: 99%

“…In this atmosphere, the OECD developed a standard test method, and in 2016, the Comet assay method using experimental animals was approved, and it is proposed as a final screening data to confirm carcinogenicity along with the micronucleus test [21,22]. Although in vitro testing has not yet been approved as an OECD Test Guideline, research for standardization is continuing because it has advantages in genotoxicity studies on nanomaterials [23][24][25][26].…”

Section: Introductionmentioning

confidence: 99%

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Genotoxicity study of 2-methoxyethanol and benzalkonium chloride through Comet assay using 3D cultured HepG2 cells

Awe

Sangodoyin

Ogundiran

2022

Environ Anal Health Toxicol

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The aim of the study was to assess the occurrence and distribution of organophosphate compounds residue in soil, surface water, sediment, and banana crops in Araromi farm settlement, Osun State, Nigeria. Organophosphate pesticide residues were determined using a gas chromatography equipped with Flame-Ionization Detection (GC-FID) in 16 soil samples from cocoa and banana farms, 6 water and sediment samples each, and 8 banana samples from 4 farms in the study site. Fourteen organophosphate compounds were detected (acephate, omethoate, dementon-s-methyl, dimethoate, tolcofos-methyl, pirimiphos-methyl, malathion, chlorpyrifos, methidathion, prothiofos, profenofos, ethion, azinphos-methyl and pyrazophos). Tolclofos-methyl, pirimiphos-methyl and prothiofos were detected in all the soil and sediment samples with concentration ranges of 1.9-12.9, 2.25-6.98 and 3.38-9.89 mg/kg respectively in soil and 8.13-9.83, 2.82-25.1 and 3.70-19.5 mg/kg respectively in sediment. Dimethoate, pirimiphos-methyl and prothiofos with concentration ranges, 0.06-0.28, 0.09-0.18 and 0.16-6.11 mg/L respectively were mostly detected in water samples while dimethoate, tolcofos-methyl, malathion, methidathion, prothiofos, ethion and azinphos-methyl compounds were detected in all the banana samples with concentration ranges, 3.40-12.0, 1.82-6.26, 5.73-9.48, 29.7-145, 8.24-20.1, 3.87-9.35 and 3.66-12.2 mg/kg respectively. The organophosphate mean residue concentrations were mostly significantly higher than the Maximum Residue Limits (MRL) at p<0.05. Across the three samples, only pirimiphos-methyl was significantly higher in water samples, omethoate in sediment; acephate, dementon-s-methyl and chlorpyrifos in banana were also not significantly higher at p<0.05. A strong positive significant correlation was observed between the organophosphate compounds in the banana and water samples (R=0.77, p=0.002) at p<0.05. The occurrence of organophosphate compounds in concentrations above MRLs may pose serious environmental and health risks.

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Quantitative comparison of in vitro genotoxicity between metabolically competent HepaRG cells and HepG2 cells using the high-throughput high-content CometChip assay

Seo

Tryndyak

et al. 2019

Arch Toxicol

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Assessment of the predictive capacity of the optimized in vitro comet assay using HepG2 cells

Cited by 14 publications

References 40 publications

An overview of apoptosis assays detecting DNA fragmentation

An overview of apoptosis assays detecting DNA fragmentation

Genotoxicity study of 2-methoxyethanol and benzalkonium chloride through Comet assay using 3D cultured HepG2 cells

Quantitative comparison of in vitro genotoxicity between metabolically competent HepaRG cells and HepG2 cells using the high-throughput high-content CometChip assay

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