Ischemia-reperfusion injuryExperimental: LiverHepatocyte Viability and Adenosine Triphosphate Content Decrease Linearly Over Time During Conventional Cold Storage of Rat Liver Grafts
Section snippets
Procurement of Rat Livers
Experiments were performed using female Lewis rats (n = 30) weighing 150 to 200 g (Charles River Labs, Wilmington, Mass). The animals were maintained in accordance with National Research Council guidelines and the experimental protocols were approved by the Subcommittee on Research Animal Care, Massachusetts General Hospital. Subjects were anesthetized with isoflurane (Forane, Baxter, Deerfield, IL) using a Tech 4 vaporizer (Surgivet, Waukesha, Wis). The liver was harvested using the technique
Cell Yield During SCS
To assess the viability of each graft, the live hepatocyte fraction was determined at various SCS storage times. Figure 1A shows the results of the hepatocyte isolations for each interval. Cell yields decreased consistently with the increase in storage time; the 120-hour control interval showed a yield of 0.87 ± 0.42 × 106 hepatocytes per gram of liver tissue, or 3.3% of the amount of freshly isolated hepatocytes (25.95 × 106 cells/g liver). Linear regression analysis revealed a correlation
Discussion
In this study we have shown that when a healthy liver is stored ex vivo there is a strong association between the amount of viable hepatocytes and the ATP level, both of which decrease linearly with time. The dynamic correlation of this process supports further exploration into the use of cellular energy levels as a quantitative criterion of graft viability. For example, previous studies have shown that rat livers can be stored and transplanted after an average SCS time of 18 to 24 hours15, 16,
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Mitochondrial energy synthesis during cold preservation and after reperfusion in liver transplantation [Translation]
Nippon Geka Gakkai Zasshi
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Tim Berendsen was funded by the Professor Michael-van Vloten Foundation, National Institutes of Health (R01 DK59766, R01 EB 008678, R00 DK080942), Shriners Hospitals for Children.