Protection of liver sinusoidal endothelial cells using different preservation solutions.

Abstract

Objective: Donor liver preservation methods and solutions have evolved over the last years. Liver sinusoidal endothelial cell (LSEC) barrier function and integrity during preservation is crucial for outcomes of liver transplantation. Therefore, the present study aimed to determine optimal preservation of LSEC barrier function and integrity, using different preservation solutions.

Methods: Human Umbilical Vein Endothelial Cells (HUVEC) and LSEC were incubated in either University of Wisconsin machine perfusion solution (UW-MPS), histidine-tryptophan-ketoglutarate (HTK), or endothelial cell growth medium 2 (EGM2) (as a golden standard for cell culturing). Endothelial integrity was assessed by measurement of cellular morphology and expression of membrane proteins ; PECAM-1, ICAM-1, and Fc-gamma receptor CD32b (FcΥRCD32b). Endothelial barrier function was measured by electric cell-substrate impedance sensing (ECIS). Cellular respondence to inflammatory stimuli with tumor necrosis factor-alpha (TNF-α), was tested by studying trans-endothelial migration (TEM) under flow conditions. Differences in these parameters were analyzed between the different preservation solutions.

Results: PECAM-1 expression was high for all preservation solutions in HUVEC and LSEC. ICAM-1 expression was increased in both LSEC and HUVEC in all preservation solutions plus TNF-α. UW reduced PECAM-1 expression, whereas EGM2 medium promoted barrier function in LSEC and HUVEC, and monolayer recovery after wounding was best achieved in cells incubated in EGM2. LSEC and HUVEC incubated with EGM2 plus TNF-α both supported neutrophil adhesion and TEM, but much less to none when incubated in UW plus TNF-α.

Conclusions: Overall, EGM2 showed the best results in preserving endothelial barrier function for both HUVEC and LSEC.