HISTOLOGICAL AND BIOCHEMICAL EVALUATION OF PRECISION-CUT LIVER SLICES

Author： Fisher Robyn L. Ulreich Judith B. Nakazato Paul Z. Brendel Klaus

ISSN： 1537-6524

Source： Toxicology Mechanisms and Methods, Vol.11, Iss.2, 2001-04, pp. : 59-79

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Abstract

Human and animal precision-cut tissue slices are being widely used to obtain drug metabolism and toxicity profiles in vitro. These data are then used to predict what might be seen in human patients. The accuracy of the extrapolation of findings based on human or animal in vitro systems to the findings that occur in vivo is dependent on the quality of the in vitro system. This study investigates the optimal thickness of tissue slices and the optimal incubation system, as determined by histological evaluation (light and electron microscopy) and two biochemical parameters (adenosine triphosphate [ATP] content and potassium [K⁺] retention). The three incubation systems tested were the dynamic organ culture system and the roller incubation system for volatiles, which are surface culture systems, and the multiwell plate culture system, which is a submersion culture system. The cellular glycogen content of canine liver slices was determined using slices fixed in 10% buffered formalin and a periodic-acid Schiff stain. After determining the optimal slice thickness (≈ 150 μm) and incubation system (the dynamic organ culture system) using gross histological evaluation, the liver slices were incubated for 6, 12, 24, 48, and 72 h, and the slices were evaluated using electron microscopy. The micrographs showed that the hepatocytes displayed normal morphology for up to 72 h. These histological findings were compared to the biochemical findings. ATP content was found to be more sensitive in detecting cellular degeneration than K⁺ retention but less sensitive than the histological evaluation. The use of tissue slices that are produced and incubated under optimal conditions can be a reliable in vitro method of investigating morphologic and metabolic alterations produced by a variety of agents, including drugs and toxicants.