Serum levels of AST as markers

of hepatocyte injury were measured 2 days after transplantation. Mice were prepared 1 hour after transplantation for intravital fluorescence microscopy as described16 on a Leica CLS 150× microscope. Microscopy sequences were captured by a camera and recorded by a video system for offline evaluation. Total RNA was extracted from liver tissue using TRIzol reagent. Quantitative real-time polymerase chain Metformin mouse reaction amplification and data analysis were performed using an ABI Prism 7000 Sequence Detection System. Results were quantified as fold induction comparison with baseline after normalization to 18S RNA. Liver tissue was prepared for scanning electron microscopy 3

hours after transplantation: the graft was flushed with 3% polyvinylpyrrolidone in Hank’s balanced salt solution. The fixed liver tissues were cut into small pieces. The specimen were then washed with phosphate-buffered saline and stored at 4°C until further processing for scanning electron microscopy. Values are expressed as the mean ± SD. The data were analyzed using GraphPad Prism version 5 software. Differences between groups were evaluated using an unpaired t test. Differences Regorafenib in vitro were considered statistically significant at P < 0.05. To evaluate whether serotonin had an effect on SFS OLT and could improve liver regeneration, we performed 30% OLT. The recipient find more was treated with the serotonin agonist DOI or with saline, until grafts were harvested 48 hours after surgery. Ki-67 and PCNA staining were analyzed on liver sections by immunohistochemistry (Fig. 1A,B). Both showed significantly enhanced hepatocyte proliferation in DOI-treated

liver grafts when compared with control grafts. Fig. 1C,E (control) and Fig. 1D,F (DOI) demonstrate a strong induction of proliferation markers by DOI. Ischemia/reperfusion injury is inevitable in organ transplantation.17, 18 It may be particularly harmful and exacerbate the loss of function in liver grafts contributing to SFS syndrome.19 To test the impact of DOI on ischemia/reperfusion injury of SFS liver grafts, serum AST was tested 2 days after 30% OLT. AST levels were elevated in the recipient control group, whereas application of DOI significantly blunted tissue injury in recipients (Fig. 1G) (P = 0.027). Hematoxylin-eosin staining of embedded liver graft tissue disclosed diffuse microvesicular steatosis in the control (Fig. 1H). Few neutrophils and rare small foci of necrosis were present in control animals. SECs are highly susceptible to cold ischemic injury.18 Preservation of intact SECs is key for a successful OLT.20 In our model of partial OLT, all grafts are inherently exposed to a short period of cold ischemia. Therefore, we studied SEC on hematoxylin-eosin–stained biopsies and by scanning electron microscopy 3 hours after reperfusion.