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Determination of Toxicity
Published in David Woolley, Adam Woolley, Practical Toxicology, 2017
Of the systems available for studying effects in the liver, isolated single cells have been the most popular. However, as indicated above, they have disadvantages in that they lose architectural polarity and undergo significant changes in regulation. In addition, they rapidly lose metabolic competence so that their viability is limited. Although subcellular systems are used, including purified enzymes and microsomal preparations, as we will see in the section on metabolism below, hepatocytes have the advantage that they are a fully functional cell complete with membranes and intracellular relationships. Another subcellular system that is regularly used is S9 mix, derived from the livers of rats, treated with an enzyme inducer, and used in genotoxicity studies where metabolic activation of the test substance is necessary. Precision-cut tissue slices are increasingly used as an evolution of the initial more crudely cut slices; they are the basis from which 3-D models and bioreactors have evolved.
Comparative study of nanoparticle uptake and impact in murine lung, liver and kidney tissue slices
Published in Nanotoxicology, 2020
Roberta Bartucci, Abhimata Paramanandana, Ykelien L. Boersma, Peter Olinga, Anna Salvati
Precision-cut tissue slices were prepared as described earlier (de Graaf et al. 2010). Briefly, liver and lung cores were prepared by using biopsy punches with a 5 mm diameter cylindrical tip and afterwards cut into slices with Krumdieck Tissue Slicer MD6000 (Alabama R&D, Munford, TN, USA). Due to their small size, the kidneys were positioned entirely in the core holder of the Krumdieck tissue slicer. The slicer was filled with ice-cold Krebs–Henseleit buffer supplemented with 25 mM d-glucose (Merck, Darmstadt, Germany), 25 mM NaHCO3 (Merck, Darmstadt, Germany), and 10 mM Hepes (MP Biomedicals GmbH, Germany), and saturated with a mixture of 95% oxygen and 5% CO2. Liver and kidney slices were about 200–250 µm thick and their weight was of around 5 mg, whereas the lung slices were 250–300 µmg thick and their weight (which includes the weight of the agarose in the section) was of roughly 6 mg. After the slicing procedure, precision-cut tissue slices prepared from the three organs were preserved in University of Wisconsin organ preservation solution (UW) (DuPont Critical Care, Waukegan, IL, USA) on ice until further use.