CYTOTOXICITY OF RETINOIC ACID, MENADIONE AND AFLATOXIN B-1 IN RAT-LIVER SLICES USING NETWELL INSERTS AS A NEW CULTURE SYSTEM

Precision-cut rat liver slices were used to develop a new dynamic incu bation system in which histomorphology and measurement of the release of lactate dehydrogenase (LDH) and the conversion of MTT were applied to evaluate cytotoxicity. Liver slices, precision-cut using a Krumdiec k tissue slicer, were cultured in a new system using 200-mu m polyeste r mesh Netwell inserts in six-well cell-culture clusters on a rocker p latform at 37 degrees C and 40% O-2. The major advantage of this new c ulture system is the easy way in which slices can be manipulated and t he culture medium be sampled or changed. Rat liver slices were exposed for 4 hr to retinoic acid (RA), menadione or aflatoxin B-1 (AFB(1)). Directly after treatment and after an additional 20-hr recovery period , histomorphological observations of slices were made, and LDH release and MTT conversion were measured. Slices exposed to RA showed dose-re lated cytotoxicity in the MTT assay only. The cytotoxic response to AF B(1) was more pronounced in the assay of LDH release than in the MTT a ssay. Histomorphology, LDH release and the MTT assay revealed cytotoxi c effects induced by menadione. We conclude that culturing liver slice s using Netwell inserts is a good alternative to other culture systems for testing non-volatile compounds.