Human liver stem cell‐derived extracellular vesicles reduce injury in a model of normothermic machine perfusion of rat livers previously exposed to a prolonged warm ischemia

AbstractHepatic ischemia-reperfusion injury (IRI) is observed in liver transplantation and hepato-biliary surgery and is associated with an inflammatory response. Human liver stem cell-derived extracellular vesicles (HLSC-EV) have been demonstrated to reduce liver damage in different experimental settings by accelerating regeneration and by modulating inflammation. The aim of the present study was to investigate whether HLSC-EV may protect liver from IRI in a mouse experimental model. Segmental IRI was obtained by selective clamping of intrahepatic pedicles for 90 min followed by 6 h of reperfusion. HLSC-EV were administered intravenously at the end of the ischemic period and histopathological and biochemical alterations were evaluated in comparison with controls injected with vehicle alone. Intra liver localization of labeled HLSC-EV was assessed by in in vivo Imaging System (IVIS) and the internalization into hepatocytes was confirmed by fluorescence analyses. As compared to the control group, administration of 3 × 109 particles (EV1 group) significantly reduced alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) release, necrosis extension and cytokines expression (TNF-α, CCL-2 and CXCL-10). However, the administration of an increased dose of HLSC-EV (7.5 × 109 particles, EV2 group) showed no significant improvement in respect to controls at enzyme and histology levels, despite a significantly lower cytokine expression. In conclusion, this study demonstrated that 3 × 109 HLSC-EV were able to modulate hepatic IRI by preserving tissue integrity and by reducing transaminases release and inflammatory cytokines expression. By contrast, a higher dose was ineffective suggesting a restricted window of biological activity.

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First human liver transplantation using a marginal allograft resuscitated by normothermic machine perfusion

Liver Transplantation ◽

10.1002/lt.24369 ◽

2015 ◽

Vol 22 (1) ◽

pp. 120-124 ◽

Cited By ~ 59

Author(s):

Thamara Perera ◽

Hynek Mergental ◽

Barney Stephenson ◽

Garrett R. Roll ◽

Hentie Cilliers ◽

...

Keyword(s):

Liver Transplantation ◽

Human Liver ◽

Machine Perfusion ◽

Normothermic Machine Perfusion

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The effect of blood cells retained in rat livers during static cold storage on viability outcomes during normothermic machine perfusion

Scientific Reports ◽

10.1038/s41598-021-02417-6 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Omar Haque ◽

Casie A. Pendexter ◽

Benjamin T. Wilks ◽

Ehab O. A. Hafiz ◽

James F. Markmann ◽

...

Keyword(s):

Cold Storage ◽

Blood Cells ◽

Energy Charge ◽

Ischemic Injury ◽

Machine Perfusion ◽

University Of Wisconsin ◽

Normothermic Machine Perfusion ◽

Preservation Solutions ◽

Positive Correlations ◽

Rat Livers

AbstractIn transplantation, livers are transported to recipients using static cold storage (SCS), whereby livers are exposed to cold ischemic injury that contribute to post-transplant risk factors. We hypothesized that flushing organs during procurement with cold preservation solutions could influence the number of donor blood cells retained in the allograft thereby exacerbating cold ischemic injury. We present the results of rat livers that underwent 24 h SCS after being flushed with a cold University of Wisconsin (UW) solution versus room temperature (RT) lactated ringers (LR) solution. These results were compared to livers that were not flushed prior to SCS and thoroughly flushed livers without SCS. We used viability and injury metrics collected during normothermic machine perfusion (NMP) and the number of retained peripheral cells (RPCs) measured by histology to compare outcomes. Compared to the cold UW flush group, livers flushed with RT LR had lower resistance, lactate, AST, and ALT at 6 h of NMP. The number of RPCs also had significant positive correlations with resistance, lactate, and potassium levels and a negative correlation with energy charge. In conclusion, livers exposed to cold UW flush prior to SCS appear to perform worse during NMP, compared to RT LR flush.

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Cold Preflush of Porcine Kidney Grafts Prior to Normothermic Machine Perfusion Aggravates Ischemia Reperfusion Injury

Scientific Reports ◽

10.1038/s41598-019-50101-7 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Gregor Fabry ◽

Benedict M. Doorschodt ◽

Tim Grzanna ◽

Peter Boor ◽

Aaron Elliott ◽

...

Keyword(s):

Reperfusion Injury ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Warm Ischemia ◽

Total Output ◽

Machine Perfusion ◽

Buffer Solution ◽

Preservation Method ◽

Fractional Excretion Of Sodium ◽

Normothermic Machine Perfusion

Abstract Normothermic machine perfusion (NMP) of kidney grafts is a promising new preservation method to improve graft quality and clinical outcome. Routinely, kidneys are washed out of blood remnants and cooled using organ preservation solutions prior to NMP. Here we assessed the effect of cold preflush compared to direct NMP. After 30 min of warm ischemia, porcine kidneys were either preflushed with cold histidine-tryptophan-ketoglutarate solution (PFNMP group) prior to NMP or directly subjected to NMP (DNMP group) using a blood/buffer solution. NMP was performed at a perfusion pressure of 75 mmHg for 6 h. Functional parameters were assessed as well as histopathological and biochemical analyses. Renal function as expressed by creatinine clearance, fractional excretion of sodium and total output of urine was inferior in PFNMP. Urine protein and neutrophil gelatinase-associated lipocalin (NGAL) concentrations as markers for kidney damage were significantly higher in the PFNMP group. Additionally, increased osmotic nephropathy was found after PFNMP. This study demonstrated that cold preflush prior to NMP aggravates ischemia reperfusion injury in comparison to direct NMP of warm ischemia-damaged kidney grafts. With increasing use of NMP systems for kidneys and other organs, further research into graft flushing during retrieval is warranted.

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The Delivery of Stem Cell Therapy to Extended Criteria Donor Human Livers Using Normothermic Machine Perfusion

10.26226/morressier.58eb975cd462b80290b4fab2 ◽

2017 ◽

Author(s):

Richard Wallace Laing

Keyword(s):

Stem Cell ◽

Cell Therapy ◽

Stem Cell Therapy ◽

Machine Perfusion ◽

Extended Criteria Donor ◽

Normothermic Machine Perfusion ◽

Human Livers

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Extracellular vesicles from human liver stem cells inhibit renal cancer stem cell‐derived tumor growth in vitro and in vivo

International Journal of Cancer ◽

10.1002/ijc.32925 ◽

2020 ◽

Vol 147 (6) ◽

pp. 1694-1706 ◽

Cited By ~ 4

Author(s):

Alessia Brossa ◽

Valentina Fonsato ◽

Cristina Grange ◽

Stefania Tritta ◽

Marta Tapparo ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cancer Stem Cell ◽

Tumor Growth ◽

Extracellular Vesicles ◽

Renal Cancer ◽

Human Liver ◽

Liver Stem Cells

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Human Liver Stem Cell Derived Extracellular Vesicles Alleviate Kidney Fibrosis by Interfering with the β-Catenin Pathway through miR29b

International Journal of Molecular Sciences ◽

10.3390/ijms221910780 ◽

2021 ◽

Vol 22 (19) ◽

pp. 10780

Author(s):

Sharad Kholia ◽

Maria Beatriz Herrera Sanchez ◽

Maria Chiara Deregibus ◽

Marco Sassoè-Pognetto ◽

Giovanni Camussi ◽

...

Keyword(s):

Stem Cell ◽

Extracellular Vesicles ◽

Human Liver ◽

Kidney Injury ◽

Loss Of Function ◽

Kidney Fibrosis ◽

Clinical Models ◽

Collagen Type 1

Human liver stem-cell-derived extracellular vesicles (HLSC-EVs) exhibit therapeutic properties in various pre-clinical models of kidney injury. We previously reported an overall improvement in kidney function following treatment with HLSC-EVs in a model of aristolochic acid nephropathy (AAN). Here, we provide evidence that HLSC-EVs exert anti-fibrotic effects by interfering with β-catenin signalling. A mouse model of AAN and an in vitro pro-fibrotic model were used. The β-catenin mRNA and protein expression, together with the pro-fibrotic markers α-SMA and collagen 1, were evaluated in vivo and in vitro following treatment with HLSC-EVs. Expression and functional analysis of miR29b was performed in vitro following HLSC-EV treatments through loss-of-function experiments. Results showed that expression of β-catenin was amplified both in vivo and in vitro, and β-catenin gene silencing in fibroblasts prevented AA-induced up-regulation of pro-fibrotic genes, revealing that β-catenin is an important factor in fibroblast activation. Treatment with HLSC-EVs caused increased expression of miR29b, which was significantly inhibited in the presence of α-amanitin. The suppression of the miR29b function with a selective inhibitor abolished the anti-fibrotic effects of HLSC-EVs, resulting in the up-regulation of β-catenin and pro-fibrotic α-Sma and collagen type 1 genes. Together, these data suggest a novel HLSC-EV-dependent regulatory mechanism in which β-catenin is down regulated by HLSC-EVs-induced miR29b expression.

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Experimental perfusion device for preservation of donor’s liver

Grekov s Bulletin of Surgery ◽

10.24884/0042-4625-2017-176-3-88-92 ◽

2017 ◽

Vol 176 (3) ◽

pp. 88-92

Author(s):

S. F. Bagnenko ◽

O. N. Reznik ◽

A. E. Skvortsov ◽

A. V. Lopota ◽

N. A. Gryaznov ◽

...

Keyword(s):

Biochemical Parameters ◽

Warm Ischemia ◽

Central Research ◽

Machine Perfusion ◽

Clinical Tests ◽

Donor Liver ◽

Porcine Liver ◽

Design Institute ◽

Normothermic Perfusion ◽

Normothermic Machine Perfusion

OBJECTIVE. The authors aimed to develop complex perfusion medical device in order to successfully maintain viability of donor liver until transplantation. MATERIAL AND METHODS. Experimental perfusion device for normothermic perfusion donor’s liver was developed and tested in the Central Research and Experimental-design Institute of Robotics and Technical Cybernetics. RESULTS. The pre-clinical tests were performed on porcine liver. It was demonstrated that normothermic machine perfusion could restore liver function after 30 minutes of warm ischemia time and provide normalization of biochemical parameters of organ functioning.

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