Development of an intact hepatocyte activation system for routine use with the mouse lymphoma assay.
We have developed a method for cocultivating primary rat hepatocytes with L5178Y/TK+/- -3.7.2C mouse lymphoma cells. This method should provide a means of stimulating more closely in-vivo metabolism compared to metabolism by liver homogenates, while still being useful for routine screening. Hepatocytes were isolated from 200-250 gm adult male Sprague-Dawley rats; 1 X 10(6) viable hepatocytes were seeded per flask. Rapid attachment of the hepatocytes (2 hr) was obtained by using fibronectin-coated 25-cm2 tissue culture flasks. Cocultivated cultures were incubated at 37 degrees C on a platform rocker at 32 oscillations per minute. A 16-hr cocultivated period was selected. With this hepatocyte activation methodology, CP, DMN, DMBA, and B(a)P, genotoxins that require metabolic activation, could be detected as mutagens in L5178Y/TK+/- cells.
D000818:Animals; D001564:Benzo(a)pyrene / Q000378:metabolism / Q000633:toxicity; D001711:Biotransformation; D002455:Cell Division; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003520:Cyclophosphamide / Q000378:metabolism / Q000633:toxicity; D008099:Liver / Q000166:cytology; D008223:Lymphoma / Q000235:genetics*; D051379:Mice; D008862:Microsomes, Liver / Q000378:metabolism*; D009152:Mutagenicity Tests / Q000379:methods*; D009154:Mutation / Q000187:drug effects*; D051381:Rats
D001564:Benzo(a)pyrene; D003520:Cyclophosphamide