The Liverbeads as a tool for the comet assay.
Liverbeads, cryopreserved hepatocytes entrapped within an alginate matrix, were examined for their relevance in the comet assay. It was estimated by their capacity to activate the indirectly acting mutagens, cyclophosphamide (CP), benzo[a]pyrene (BP), dimethylbenzanthracene (DMBA) and 2-acetylaminofluorene (2-AAF), into DNA reactive metabolites. The comet assay performed in alkaline condition is a sensitive method for detecting strand breaks at the level of individual cells and allows use of quiescent cells. Experimental conditions as treatment time, cell density, beads dissociation and viability were investigated. Significant statistical positive results assessed by the tail extent moment (TEM) were observed with both human and rat Liverbeads after 12h duration incubation compared to metabolic non-competent cells, HeLa S3. Due to the maintenance of specific functions assessed by the observed capacity to metabolize xenobiotics, Liverbeads represent a suitable tool system, easy to handle, for the detection of promutagens using the comet assay.
Vian
Laurence
L
Laboratoire de Toxicologie, Faculté de Pharmacie, BP 14491, 34093 Montpellier Cedex 5, France. lvian@pharma.univ-montp1.fr
Yusuf
Ayda
A
Guyomard
Claire
C
Cano
Jean-Paul
JP
D015073:2-Acetylaminofluorene / Q000009:adverse effects; D015127:9,10-Dimethyl-1,2-benzanthracene / Q000009:adverse effects; D000368:Aged; D000818:Animals; D001564:Benzo(a)pyrene / Q000009:adverse effects; D001711:Biotransformation; D002273:Carcinogens / Q000009:adverse effects; D020552:Comet Assay; D003520:Cyclophosphamide / Q000009:adverse effects; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000187:drug effects*; D005260:Female; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008297:Male; D008863:Microspheres; D008875:Middle Aged; D009153:Mutagens / Q000009:adverse effects*; D011355:Prodrugs / Q000009:adverse effects*; D051381:Rats; D012680:Sensitivity and Specificity
D002273:Carcinogens; D004273:DNA, Neoplasm; D009153:Mutagens; D011355:Prodrugs; D001564:Benzo(a)pyrene; D015127:9,10-Dimethyl-1,2-benzanthracene; D003520:Cyclophosphamide; D015073:2-Acetylaminofluorene