hepatocyte cultures

/Tag:hepatocyte cultures

A Preliminary Comparison of LiverBeadsTM with a Conventional Rat Hepatocyte Culture Preparation: Some Aspects of Xenobiotic Metabolism and Related Toxicity

Alison H. Hammond, Michael J. Garle and Jeffrey R. Fry

LiverBeadsTM are made of hepatocytes that are immobilised and cryopreserved in alginate gel. They have great potential as an easily transportable and easily handled source of hepatocytes for use in in vitro pharmacotoxicology. In this study, we compared the drug metabolising capacity of LiverBeads in culture with that of conventional cultures and of cultures derived from cryopreserved cells. Trypan blue exclusion, lactate dehydrogenase and DNA content were measured in LiverBead cultures. The levels were all similar to those of the conventional cultures, as were the toxicities of precocene II and allyl alcohol, although more variability was seen in the LiverBeads than in the conventional cultures. The cytochrome P450-dependent activity 3,4-dimethyl-7-ethoxycoumarin-O-dealkylase, was reduced in the LiverBeads when compared to the conventional cultures, although the pattern of conjugation was very similar. In addition, the inducibility of cytochrome P4504A was demonstrated in LiverBeads. Cultures from cryopreserved cells were more susceptible to the toxicants tested, and contained less lactate dehydrogenase and DNA than the conventional cultures. Overall, in terms of the aspects of drug metabolism measured, the cultures from LiverBeads appeared to be equivalent to conventional cultures, and superior to cultures from cryopreserved cells.
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Comparison of Hepatocyte Cultures and Liver Slices in In Vitro Toxicity Testing

Elisabeth George, Janice Murdock, Mike Aylott and Carl Westmoreland

The aim of this study was to compare the in vitro toxicities of two hepatotoxins in hepatocyte cultures and in liver slices from both rats and dogs. Hepatocytes and liver slices were pre-incubated for 2 hours and then exposed to galactosamine or paracetamol, both of which mainly induce liver necrosis in vivo. Following exposure to the compounds for 20 hours, neutral red uptake (NRU [hepatocyte cultures only]), MTT reduction, and reduced glutathione (GSH), adenosine triphosphate (ATP) and protein content, were used to measure the toxicity induced. In general, galactosamine and paracetamol exposure caused comparable levels of toxicity in hepatocyte cultures and in liver slices. For galactosamine, no consistent differences were seen between hepatocyte cultures and liver slices. With paracetamol, the toxic effects were generally slightly more pronounced in hepatocyte cultures than in liver slices, and the preparations from dog liver were more sensitive than those from rat liver to paracetamol exposure. These results are in agreement with previously described species differences in vitro. NRU and GSH content were more sensitive and more consistent endpoints than MTT reduction, ATP content or protein content. Liver slices appeared to lose viability over the 20 hours in culture. Therefore, it can be concluded that liver slices should only be used in relatively short-term investigations.
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