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Use of primary rat and human hepatocyte sandwich cultures for activation of indirect carcinogens: Monitoring of DNA strand breaks and gene mutations in co-cultured cells

: Fahrig, R.; Rupp, M.; Steinkamp-Zucht, A.; Bader, A.


Toxicology in vitro 12 (1998), S.431-444
ISSN: 0887-2333
Fraunhofer ITA ( ITEM) ()
benzo(a)pyrene; Carcinogens; cell culture; Dimethylbenzanthracene; DNA; genetic toxicology; Genotoxicology; Hepatocytes; Human cell culture; mutation; rat

Loss of cytochrome P-450 content is a common feature in conventional culture systems of primary hepatocytes. In contrast to the standard in vitro situation, in vivo each hepatocyte is exposed to an extracellular matrix (space of Disse) at two opposing basolateral surfaces. This in vivo symmetrie has been reconstructed in vitro by culturing rat or human hepatocytes within two layers of collagen, thus forming a sandwich configuration. Activation of dimethylbenzanthracene (DMBA) or benzol(a)pyrene (BaP) was studied in rat and human hepatocytes. Genotoxic effects were studied in a three-dimensional co-culture model between sandwich hepatocytes and mammalian cells using the comet assay for detection of DNA strand breaks.