Dynamic Organ Culture is Superior to Multiwell Plate Culture for Maintaining Precision-Cut Tissue Slices: Optimization of Tissue Slice Culture, Part 1

Abstract

The increase in the use of precision-cut tissue slices for metabolism and toxicity studies has resulted in the development of various incubation systems and techniques for culturing the slices. This has led to inconsistencies in data obtained from different laboratories. For data to be comparable from one laboratory to another, reliable and consistent incubation systems must be used that will give the researcher the most optimal tissue slice viabilities. This study compares and contrasts the dynamic organ culture system (surface culture) and the multiwell plate culture system (submersion culture). Rat liver slices were produced using the Brendel/Vitron tissue slicer under oxygenated and ice-cold V-7 preservation solution. The slices (200 μm thick) were incubated in Waymouth's medium + 10% fetal calf serum (1.7 mL) containing either sodium bicarbonate or Hepes and gassed with either 95%/5% O2/CO2 or 95%/5% air/CO2. Slice viability was assessed at 6, 24, 48, and 72 h using ATP content, K+ retention, protein synthesis, alanine aminotransferase and lactate dehydrogenase leakage, and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) reduction. 7-Ethoxycoumarin metabolism was also used to assess the xenobiotic metabolic activity of liver slices. The results indicated that the dynamic organ culture system maintained rat liver slices at a higher level of viability than the multiwell plate culture system and that Waymouth's medium gassed with 95% O2/5% CO2 was the best incubation condition for both systems. Thus, it is essential to optimize slice viability in order to obtain reliable data that will ultimately be used to predict what will be seen in humans.