Adenosine 5'triphosphate transport and accumulation during the cold preservation of rat hepatocytes in University of Wisconsin solution

Figure 1 A: Effect of the addition of ATP to the UW solution on intracellular ATP content in hepatocytes. Cells were cold stored (0 °C) up to 72 h in UW solution containing ATP: 5 mmol/L (b-group), 10 mmol/L (c-group) and without ATP (a-group). ATP was determined at time 0 and 72 h. Data are given as the mean±SD for hepatocytes prepared from three rats for each group. The asterisk indicates that after 72 h of cold storage, the ATP content of hepatocytes from groups a and b is statistically different from c-group at ^aP<0.05; B: ATP content of freshly isolated and cold-stored hepatocytes during rewarming. Cells from a-, b- and c-groups were cold preserved (0 °C, 72 h); they were later resuspended and incubated at 37 °C for 120 min in KHR. ATP was determined at time 0 and 120 min. Data are expressed as the mean±SD for three separate preparations. At time 0, the asterisk indicated that groups a and b are different from groups c and controls (^aP<0.05); at 120 min, the asterisk indicated that groups a and b are different from groups c and controls (^aP<0.05).

Figure 2 Time course of [³H]-ATP (10 mmol/L) (4. 1 μCi/mL) accumulation by cold-stored hepatocytes. Cells were incubated (0 °C) in UW solution. True uptake was the difference between total uptake and nonspecific binding to cells (as described under Materials and Methods). Results are expressed as the mean±SD from three determinations for each point.

Figure 3 [³H]-ATP kinetic in cold-stored hepatocytes. Cells preserved in UW solution (0 °C) were incubated with [³H]-ATP (3.13 μCi/mL) and varying concentrations of extracellular ATP (of 0.5, 1.0, 2.5, 5.0 and 10.0 mmol/L) for 10 min. Results are expressed as the mean±SD from four determinations for each point.

Figure 4 Time course evolution of extracellular added ATP and nucleotides over 60 min of cold storage. Hepatocytes were separated from their incubation medium after the desired period of storage and samples of medium assayed. Results are expressed as the mean±SD from four experiments. The white bar ATP-UW represents the values obtained from UW solution+10 mmol/L ATP without cells (d-group).