DETERMINATION OF OPTIMAL INCUBATION MEDIA AND SUITABLE SLICE DIAMETERS IN PRECISION-CUT LIVER SLICES - OPTIMIZATION OF TISSUE SLICE CULTURE.2.

To validate tissue slice methodology in experiments from one laborator y to another, it is important to optimize slice viability. The effects of different culture media and slice diameters were investigated in t his study. Rat and mouse liver slices (200 mu m thick) were placed in either Krebs/Hepes, Krebs/bicarbonate, Waymouth's/Hepes, or Waymouth's /bicarbonate to assess the effects of nutrient-enriched and nutrient-d eprived media as well as the effects of a Hepes- or bicarbonate-based medium on slice viability. Overall, an enriched and bicarbonate-based medium maintained slice viability better. Determination of suitable sl ice diameters was achieved by incubating 6- and 8-mm-diameter rat, mou se, and dog liver slices in Waymouth's/bicarbonate media for 24 h. For both rat and mouse liver slices, the 8-mm-diameter slices were better maintained than the 6-mm slices, whereas both diameters were equivale nt for dog liver slices. Various nutrient-enriched media were also com pared for their ability to maintain rat liver Slice viability in cultu re. Of the 15 media tested, nine maintained slice K+ levels above 80 m u mol/g wet weight for 3 days. Rat liver slice viability could be main tained for 5 days in culture when 8-mm slices were incubated in Waymou th's/bicarbonate medium in dynamic organ culture. Liver slice viabilit y was assessed by measuring K+ retention, protein synthesis, and lacta te dehydrogenase leakage. This study shows that incubation parameters do affect liver slice viability and that these parameters need to be o ptimized.