Protection of liver sinusoidal endothelial cells using different preservation solutions

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

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Main Authors: Julia H E Houtzager, Anne-Marieke van Stalborch, Charlotte Hofstee, Thomas M van Gulik, Jaap D van Buul
Format: Article
Language:English
Published: Bioscientifica 2025-02-01
Series:Vascular Biology
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Online Access:https://vb.bioscientifica.com/view/journals/vb/7/1/VB-24-0004.xml
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author Julia H E Houtzager
Anne-Marieke van Stalborch
Charlotte Hofstee
Thomas M van Gulik
Jaap D van Buul
author_facet Julia H E Houtzager
Anne-Marieke van Stalborch
Charlotte Hofstee
Thomas M van Gulik
Jaap D van Buul
author_sort Julia H E Houtzager
collection DOAJ
description Donor liver preservation methods and solutions have evolved over the last years. Liver sinusoidal endothelial cell (LSEC) barrier function and integrity during preservation are 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. Human umbilical vein endothelial cells (HUVECs) and LSECs were incubated in either University of Wisconsin machine perfusion solution (UW-MPS), histidine-tryptophan-ketoglutarate, or endothelial cell growth medium 2 (EGM2) (as a gold 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. Cellular response 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. PECAM-1 expression was high for all preservation solutions in HUVECs and LSECs. ICAM-1 expression was increased in both LSECs and HUVECs in all preservation solutions plus TNF-α. UW reduced PECAM-1 expression, whereas EGM2 medium promoted barrier function in LSECs and HUVECs, and monolayer recovery after wounding was best achieved in cells incubated in EGM2. LSECs and HUVECs incubated with EGM2 plus TNF-α both supported neutrophil adhesion and TEM, but much less to none when incubated in UW plus TNF-α. Overall, EGM2 showed the best results in preserving endothelial barrier function for both HUVECs and LSECs.
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spelling doaj-art-78888359ad4a4b7fbe36e734166085352025-02-11T12:33:50ZengBioscientificaVascular Biology2516-56582025-02-017110.1530/VB-24-00041Protection of liver sinusoidal endothelial cells using different preservation solutionsJulia H E Houtzager0Anne-Marieke van Stalborch1Charlotte Hofstee2Thomas M van Gulik3Jaap D van Buul4Department of Surgery, Amsterdam UMC, University of Amsterdam, Amsterdam, The NetherlandsMolecular Cell Biology Lab., Department Blood Cell Research, Sanquin Research and Landsteiner Laboratory, Amsterdam, The NetherlandsMolecular Cell Biology Lab., Department Blood Cell Research, Sanquin Research and Landsteiner Laboratory, Amsterdam, The NetherlandsDepartment of Surgery, Amsterdam UMC, University of Amsterdam, Amsterdam, The NetherlandsMolecular Cell Biology Lab., Department Blood Cell Research, Sanquin Research and Landsteiner Laboratory, Amsterdam, The NetherlandsDonor liver preservation methods and solutions have evolved over the last years. Liver sinusoidal endothelial cell (LSEC) barrier function and integrity during preservation are 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. Human umbilical vein endothelial cells (HUVECs) and LSECs were incubated in either University of Wisconsin machine perfusion solution (UW-MPS), histidine-tryptophan-ketoglutarate, or endothelial cell growth medium 2 (EGM2) (as a gold 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. Cellular response 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. PECAM-1 expression was high for all preservation solutions in HUVECs and LSECs. ICAM-1 expression was increased in both LSECs and HUVECs in all preservation solutions plus TNF-α. UW reduced PECAM-1 expression, whereas EGM2 medium promoted barrier function in LSECs and HUVECs, and monolayer recovery after wounding was best achieved in cells incubated in EGM2. LSECs and HUVECs incubated with EGM2 plus TNF-α both supported neutrophil adhesion and TEM, but much less to none when incubated in UW plus TNF-α. Overall, EGM2 showed the best results in preserving endothelial barrier function for both HUVECs and LSECs.https://vb.bioscientifica.com/view/journals/vb/7/1/VB-24-0004.xmlpreservationorgan transplantendotheliumsinusoidsliver
spellingShingle Julia H E Houtzager
Anne-Marieke van Stalborch
Charlotte Hofstee
Thomas M van Gulik
Jaap D van Buul
Protection of liver sinusoidal endothelial cells using different preservation solutions
Vascular Biology
preservation
organ transplant
endothelium
sinusoids
liver
title Protection of liver sinusoidal endothelial cells using different preservation solutions
title_full Protection of liver sinusoidal endothelial cells using different preservation solutions
title_fullStr Protection of liver sinusoidal endothelial cells using different preservation solutions
title_full_unstemmed Protection of liver sinusoidal endothelial cells using different preservation solutions
title_short Protection of liver sinusoidal endothelial cells using different preservation solutions
title_sort protection of liver sinusoidal endothelial cells using different preservation solutions
topic preservation
organ transplant
endothelium
sinusoids
liver
url https://vb.bioscientifica.com/view/journals/vb/7/1/VB-24-0004.xml
work_keys_str_mv AT juliahehoutzager protectionofliversinusoidalendothelialcellsusingdifferentpreservationsolutions
AT annemariekevanstalborch protectionofliversinusoidalendothelialcellsusingdifferentpreservationsolutions
AT charlottehofstee protectionofliversinusoidalendothelialcellsusingdifferentpreservationsolutions
AT thomasmvangulik protectionofliversinusoidalendothelialcellsusingdifferentpreservationsolutions
AT jaapdvanbuul protectionofliversinusoidalendothelialcellsusingdifferentpreservationsolutions