[Research on the comet assay in vitro by plating HepG2 cells in 96-well plates].
OBJECTIVE
To provide the proof for the high throughput screening method in vitro by establishing the comet assay in vitro using HepG2 cells plated in 96-well plates.


METHODS
Plating HepG2 cells in 96-well plates, and then detecting the cytotoxicity using the neutral red method before comet assay.


RESULTS
8 genotoxicial agents could be detected using the method and the sensitivity was 100% , the ratio of the comet cells and the length of the tail of the comet were increased with the increasing dose when the cytotoxicity less than 50%. 11 non-genotoxicial agents could be detected within 12 non-genotoxicial agents and the specificity was 91%.


CONCLUSIONS
(1) This method could detect the direct genotoxic agents and the indirect genotoxic agents at the absent of the S9. (2)4-5 agents could be detected in the same 96-well plate using the method. (3) Using the method, we could obtain an dose at which the agent could cause genotoxicity. (4) By plating cells in plates in vitro, increased the speed of the detection and reduced the consumption of the agents. (5) The method could be developed into the high throughput screening method in vitro.
Wang
Zheng
Z
Department of Toxicology, Second Military Medical University, Shanghai 200433, China.
Zhang
Tianbao
T
Zhu
Yongfei
Y
Zhu
Jiangbo
J
D002273:Carcinogens / Q000633:toxicity*; D020552:Comet Assay / Q000379:methods*; D004249:DNA Damage / Q000187:drug effects; D056945:Hep G2 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009153:Mutagens / Q000633:toxicity*
D002273:Carcinogens; D009153:Mutagens