Analysis of mutagens with single cell gel electrophoresis, flow cytometry, and forward mutation assays in an isolated clone of Chinese hamster ovary cells

Abstract

We investigated the induction of DNA strand breaks in the single cell gel electrophoresis (SCGE or comet) assay with whole cell clastogenicity measured with flow cytometric analysis in cells from an isolated clone of the Chinese hamster ovary (CHO) AS52 cell line. Under identical treatment conditions the responses were compared with forward mutation at gpt using 2‐acetoxyacetylaminofluorene (2AAAF), ultraviolet radiation (UV) and ethyl methanesulfonate (EMS). Cytotoxicity for each agent was evaluated in the SCGE and forward mutation assays. Forward mutation was 4–10‐fold more sensitive than DNA strand breaks detected in the SCGE assay. For 2AAAF and EMS, the kinetics of the induction of genetic damage were similar for the three assays, although there were differences in sensitivity. With UV, the induction kinetics of gpt mutation differed from that expressed by SCGE and flow cytometric analysis. With the chemical mutagens 2AAAF and EMS, there was a high correlation between the SCGE assay and forward mutation and also between the SCGE assay and flow cytometry. There was no significant correlation between flow cytometry and forward mutation. With UV, only the SCGE assay and flow cytometry were correlated. Agent‐specific variations in the intragenomic distribution of DNA damage for each mutagen was measured in the SCGE assay. Environ. Mol. Mutagen. 32:360–368, 1998 © 1998 Wiley‐Liss, Inc.

Transformation of Iopamidol during Chlorination

“…Chinese hamster ovary (CHO) cell line AS52, clone 11−4−8 was used for the toxicity studies. 17 The CHO cells were maintained in Ham's F12 medium containing 5% fetal bovine serum, 1% antibiotics (100 U/mL sodium penicillin G, 100 μg/mL streptomycin sulfate, 0.25 μg/mL amphotericin B in 0.85% saline), and 1% glutamine at 37°C in a humidified atmosphere of 5% CO 2 . This 96-well microplate assay measures the reduction in cell density as a function of the concentration over a period of 72 h (∼3 cell cycles).…”

Comparative Mammalian Cell Toxicity of N-DBPs and C-DBPs

“…CHO cells are widely used in toxicology research; we used line AS52 clone 11-4-8 (7). This clone expresses a stable chromosome complement as well as functional p53 protein and normal DNA repair kinetics (7)(8)(9). CHO cells were maintained in Ham's F12 medium +5% fetal bovine serum (FBS) at 37°C in a humidified atmosphere of 5% C0 2 .…”

“…Drinking water disinfection by-products (DBPs) are an unintended consequence and were discovered over 30 years ago; since then over 600 DBPs have been identified (7). The vast majority of DBPs have not been chemically or biologically characterized (/, 2).…”

Hydrogen Sulfide Induces Direct Radical-Associated DNA Damage

“…For the data presented, at least eight microgels were analyzed per concentration with 25 nuclei measured per microgel. containing 5% fetal bovine serum, 1% antibiotics (100 units/mL sodium penicillin G, 100 Ag/mL streptomycin sulfate, 0.25 Ag/ mL amphotericin B, 0.85% saline), and 1% glutamine at 37jC in a humidified atmosphere of 5% CO 2 (49).…”