Liver cultures mutagenesis

Williams, G.M. 1980. The detection of chemical mutagens/carcinogens by DNA repair and mutagenesis in liver cultures In Chemical Mutagens, Principles and Methods for Their Detection vol. 6, eds. F.J. deSerres, A. 

The Detection of Chemical Mutagens/ Carcinogens by DNA Repair and Mutagenesis in Liver Cultures... 

San, R. H. C. and Williams, G. M., Rat hepatocyte primary cell culture-mediated mutagenesis of adult rat liver epithelial cells by procarcinogens. Proc. Soc. Exp. Biol. Med. 156 534-538 (1977). 

TABLE 7. Rat-Hepatocyte-Primary-Cell-Culture-Mediated Mutagenesis of Adult Rat Liver Epithelial Cells by Activation-Dependent Carcinogens"... 

In the heptatocyte-mediated mutagenesis studies, freshly isolated hepatocytes were mixed in a ratio of about 2 1 with liver epithelial cells before inoculation into culture flasks. After attachment, the two cell types were found to be well mixed and in close contact. Exposure of mixed cultures to several activation-dependent carcinogens that produce DNA repair in hepatocytes also produced mutagenesis in the cocultivated rat liver epithelial lines (Table 7) under conditions of exposure in which the carcinogens were not mutagenic to the lines in the absence of cocultivated hepatocytes. Thus, activated metabolites produced by the hepatocytes were transferred to the epithelial cells. Efforts to simplify this assay by the use of stored frozen hepatocytes are in progress. 

J. J. Berman, C. Tong, and G. M. Williams, Enhancement of mutagenesis during cell replication of cultured liver epithelial cells, Cancer Lett. 4, 277-283 (1978). 

The first cell-mediated mutagenesis system developed by Huberman and Sachs uses fibroblasts to activate the carcinogen and mutable V79 cells which are incapable of activating most carcinogens. This approach has demonstrated a correlation between polycyclic hydrocarbons and mutagenic activity of in-vivo carcinogenic activity. However, cultured fibroblasts are not able to activate certain classes of compounds, including those which cause cancer of the liver. ... 

Therefore, to increase the spectrum of compounds that can be studied in the cell-mediated mutagenesis system, Langenbach et al. have used primary cultures of rat liver cells to activate two known classes of liver carcinogens (nitrosamines and aflatoxins), and V79 cells to detect their mutagenic metabolites. A schematic diagram summarising the procedure and technique is shown in Fig. 2. Conversion from ouabain susceptibility to ouabain resistance, a mutation associated with the surface member Na /K ATPase, was used as the genetic marker. Typical data obtained with several nitrosamines and aflatoxins are summarised in Table 5. 

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