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Published on: August 4, 2012
Study of the microcirculation in hDAF transgenic rat livers xenoperfused with human blood
Silvia Tomat1, Francesco Paolo Russo, Maria Guido
1Department of Surgical and Gastroenterological Sciences, University of Padua, Via Giustiniani 2, VI piano Monoblocco, Padua, Italy.
Background:
The microcirculation was assessed in the livers of human decay accelerating factors (hDAF) and wild-type transgenic rats by fluorescent intravital microscopy, histology and histomorphology to determine the benefits of hDAF expression for the microcirculation of a rat liver xenograft perfused with human blood.
Methods:
Male hDAF transgenic rats (group A; n = 20) and wild-type Sprague-Dawley rats (group B; n = 20) were xenoperfused with human blood, while other male wild-type Sprague-Dawley rats (group C; n = 10) were perfused with allogeneic blood. Following plasma and leukocyte staining with fluorescein sodium, and platelet staining with rhodamine, the right lobe of the liver was assessed by intravital microscopy, counting the numbers of perfused sinusoids and leukocytes adhering to the endothelium per mm(2), and calculating the acinar perfusion index (Pi). The liver underwent histological assessment at the end of each experiment. Mean +/- SEM values were calculated and the Mann-Whitney U-test was used for statistical analysis.
Results:
The number of perfused sinusoids was higher in the group of hDAF rat livers (group A) and controls (group C) than in the group of non-transgenic rat livers perfused with human blood (group B) (P < 0.05), although only group C still had a significantly more perfused sinusoids than the other groups after 90 min of perfusion (P < 0.05). The acinar perfusion index was higher in groups A and C than in group B (P < 0.05); here again, only group C still had a significantly higher Pi than group B after 90 min of perfusion (P < 0.05). There was a massive accumulation of leukocytes that peaked after 5 min and persisted throughout the perfusion in all three groups. Histology showed portal and subendothelial hepatic vein hemorrhage, necrosis and inflammatory reaction, which were particularly evident in group B.
Conclusion:
In our study, rat livers transgenic for hDAF were better protected against early tissue damage by perfusion with human blood, but this did not result in a longer xenograft survival.
