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5142628 
Technical Report 
THE IN VITRO UNSCHEDULED DNA SYNTHESIS (UDS) ASSAY IN RAT PRIMARY HEPATOCYTES: VALIDATION OF IMPROVED METHODS FOR PRIMARY CULTURE INCLUDING DATA ON THE LACK OF EFFECT OF IONIZING RADIATION 
Harbach, PR; Aaron, CS; Wiser, SK; Grzegorczyk, CR; Smith, AL 
1989 
1 
Mutation Research: Environmental Mutagenesis and Related Subjects
ISSN: 0165-1161 
101 
2 
101-110 
English 
BCI:BCI198988003912 
The in vitro unscheduled DNA synthesis (UDS) assay was evaluated for inclusion in a battery of assays used at The Upjohn Company for evaluation of lead compounds in the development of new and existing drug entities. This evaluation process encompassed aspects of the isolation of hepatocytes and tests of reference mutagens and genotoxins. The flow rate of perfusion solutions and their temperatures were critical in the isolation of high viability hepatocytes in good yield. The attachment of freshly isolated hepatocytes to coverslips was greatly enhanced by coating the coverslips with type III collagen. Results of testing 12 known genotoxic agents (UV light, cyclophosphamide, 7,12-dimethylbenzanthracene, dimethyl-nitrosamine, diethylnitrosamine, 2-acetylaminofluorene, benzo[a]pyrene, methyl methanesulfonate, ethyl methanesulfonate, N-propyl-N′-nitro-N-nitrosoguanidine, benzidine and 4-aminobiphenyl) were in agreement with the literature. The use of X-ray did not induce unscheduled DNA synthesis in hepatocytes. This latter finding draws attention to the inability of this assay to detect agents which result in 'short-patch' repair of damage. © 1989. 
Hepatocytes, primary; Primary culture; Rat; Unscheduled DNA synthesis assay 
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