Development of an intact hepatocyte activation system for routine use with the mouse lymphoma assay

1 January 1987

journal article
research article
Published by Wiley in Environmental Mutagenesis

Vol. 9 (3) , 331-341
https://doi.org/10.1002/em.2860090312

Abstract

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 simulating 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 × 106 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°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.

Keywords

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