Prolonged ischemia time due to organ procurement and transport are main contributors to a decrease in organ quality, which is further enhanced during reperfusion. This so-called ischemia reperfusion injury is a main contributor to early graft dysfunction, which leads to costly and lengthy follow-up treatments or even organ loss in clinical liver transplantation. To address the ever-increasing gap between patients on waiting list and organs available for transplantations, more and more marginal grafts are transplanted.

Methods to monitor graft quality prior to transplantation are therefore highly desirable to optimize clinical outcome.

In an experimental model we used life confocal microscopy to assess murine liver graft quality. Six to ten week-old male animals were subjected to a methionine-choline-deficient (MCD) diet causing non-alcoholic fatty liver disease (NAFLD), or to the Lieber DeCarli diet producing alcohol-induced liver injury. Untreated animals served as control. In each group liver biopsies were analyzed after 45 minutes warm ischemia time (WIT) induced by liver pedicle occlusion, 24 hours cold ischemia time (CIT) respectively. All clamped livers were reperfused for 4h. Graft quality assessment was performed by measurement of serum transaminases, standard histopathology on paraffin- and cryosections and live confocal microcopy detection of inflammatory marker was conducted using PCR.

After prolonged CIT and WIT liver grafts showed a significantly decrease in cell viability when compared to naïve animals (p < 0,05) as assessed using life confocal microscopy. Animals exposed to the MCD diet showed significantly lower cell viability within the liver biopsies after CIT as well as after WIT when compared to control animals (p < 0,05). Similar results were obtained from the analysis of cell viability of animals fed with the LDC diet. Results from the life confocal microscopy were then correlated with the results from detection of serum transaminases and evaluation of standard H&E staining and results from PCR analysis. Revealing a strong correlation of cell viability, serum transaminases and tissue damage on H&E slides as well as expression of inflammatory markers within liver biopsies.

Our data demonstrate that live confocal microscopy requires minimal time for sample preparation and data acquisition and is well suitable to record organ damage prior to liver transplantation.

CITATION INFORMATION: Oberhuber R, Fodor M, Cardini B, Maglione M, Hermann M, Troppmaier J. Life Confocal Imaging of Murine Liver Biopsies as Novel Tool to Assess Graft Viability Prior to Transplantation. Am J Transplant. 2017;17 (suppl 3).

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