A sensitive serum-free colorimetric assay for the detection of cytotoxic effects of pesticides.
A new method of cell quantification in the Hybritest bioassay is described. This test is based on culturing the hybridoma cell line 1E6 in the serum-free medium RPMI-SR3 and determination of cytotoxicity by growth inhibition. By employing a tetrazolium salt (MTT), the cell number were quantified colorimetrically using an ELISA reader. This assay carried out in microtiter plates saved time, labour and expenses. It was demonstrated that the sensitivity of 1E6 to pesticide toxicity (glyphosate and cypermethrine) was considerably higher in serum-free medium than in serum-containing medium. Furthermore, it is suggested that this assay may represent an alternative to the use of living animals in toxicological experiments.
Figenschau
Y
Y
Dept. of Obstetrics and Gynaecology, University Hospital of Tromsö, Norway.
Yousef
M I
MI
Sveinbjörnsson
B
B
Bertheussen
K
K
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003124:Colorimetry; D003470:Culture Media; D004797:Enzyme-Linked Immunosorbent Assay; D005998:Glycine / Q000031:analogs & derivatives* / Q000633:toxicity; D006540:Herbicides / Q000633:toxicity*; D006825:Hybridomas / Q000166:cytology; D007306:Insecticides / Q000633:toxicity*; D007928:Lethal Dose 50; D008668:Metallothionein / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010084:Oxidation-Reduction; D011722:Pyrethrins / Q000633:toxicity*; D013056:Spectrophotometry, Ultraviolet; D013778:Tetrazolium Salts / Q000737:chemistry; D000097797:Glyphosate
D003470:Culture Media; D005998:Glycine; D006540:Herbicides; D007306:Insecticides; D008668:Metallothionein; D011722:Pyrethrins; D013778:Tetrazolium Salts; D000097797:Glyphosate; C042207:4-anisyltetrazolium blue; C017160:cypermethrin