Advancing multi-day ex vivo kidney perfusion using spatially resolved metabolomics
bioRxiv (Cold Spring Harbor Laboratory)
Abstract
ABSTRACT The ability to preserve metabolically active kidneys ex vivo for multiple days may permit reconditioning, repair and regeneration of deceased donor kidneys. However, the kidneys high metabolic demand limits its functional preservation. Current approaches focus on normothermic machine perfusion (NMP) at 37°C or hypothermic machine perfusion (HMP) at 4-8°C. At normothermia, kidneys are metabolically active but ex vivo preservation is limited to hours. During hypothermia kidneys can be preserved up to 24 hours but are metabolically inactive and suffer cold-induced injury. Therefore, we revisited sub normothermic perfusion (at 25°C) as an alternative approach to preserve human kidneys in a metabolically active state for extended periods of time. In a custom-made platform that includes a cell-free perfusate enriched with TCA cycle fuels, urine recirculation, and continuous hemofiltration we perfused discarded human kidneys up to 8 days. Using spatially resolved single cell resolution isotope tracing we demonstrate active metabolism in all the different renal cell types over this period. However, beyond 4 days cell composition of nephron segments assessed with spatial lipidomics changed substantially and injury markers such as NGAL and LDH increased in the perfusate. Up to 4 days, perfused human discarded donor kidneys maintained metabolic fluxes, functional parameters and allow for reperfusion using a porcine auto transplantation model. These data underpin that extended multi-day metabolic preservation of human kidneys is achievable using a sub normothermic perfusion platform.
Authors 19
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Marlon J.A. de Haan Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University · Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center; Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center; Leiden, the Netherlands
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center
Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center
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Franca M. R. Witjas Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine
Leiden University · Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center; Leiden, the Netherlands
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center
Leiden, the Netherlands
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Annemarie M. A. de Graaf Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine
Leiden University · Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center; Leiden, the Netherlands
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center
Leiden, the Netherlands
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Marleen E Jacobs Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University · Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center; Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center; Leiden, the Netherlands
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center
Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Center of Proteomics and Metabolomics, Leiden University Medical Center; Leiden, the Netherlands
Center of Proteomics and Metabolomics, Leiden University Medical Center
Leiden, the Netherlands
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Leiden University Medical Center
Affiliation as printed
Center of Proteomics and Metabolomics, Leiden University Medical Center; Leiden, the Netherlands
Center of Proteomics and Metabolomics, Leiden University Medical Center
Leiden, the Netherlands
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Martin A. Giera Aachen Center of Proteomics and Metabolomics Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Center of Proteomics and Metabolomics, Leiden University Medical Center; Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center; Leiden, the Netherlands
Center of Proteomics and Metabolomics, Leiden University Medical Center
Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center
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Inserm · Université de Lille · Institut Pasteur de Lille · Centre Hospitalier Universitaire de Lille
Affiliation as printed
University of Lille, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Universitaire de Lille (CHU Lille), Institute Pasteur Lille; Lille, France
Lille, France
University of Lille, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Universitaire de Lille (CHU Lille), Institute Pasteur Lille
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Inserm · Université de Lille · Institut Pasteur de Lille · Centre Hospitalier Universitaire de Lille
Affiliation as printed
University of Lille, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Universitaire de Lille (CHU Lille), Institute Pasteur Lille; Lille, France
Lille, France
University of Lille, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Universitaire de Lille (CHU Lille), Institute Pasteur Lille
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Inserm · Université de Lille · Institut Pasteur de Lille · Centre Hospitalier Universitaire de Lille
Affiliation as printed
University of Lille, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Universitaire de Lille (CHU Lille), Institute Pasteur Lille; Lille, France
Lille, France
University of Lille, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Universitaire de Lille (CHU Lille), Institute Pasteur Lille
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Inserm · Université de Lille · Institut Pasteur de Lille · Centre Hospitalier Universitaire de Lille
Affiliation as printed
University of Lille, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Universitaire de Lille (CHU Lille), Institute Pasteur Lille; Lille, France
Lille, France
University of Lille, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Universitaire de Lille (CHU Lille), Institute Pasteur Lille
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Dorottya K. de Vries Aachen Department of Surgery, Leiden University Medical Center Transplant Center
Leiden University Medical Center
Affiliation as printed
Department of Surgery, Leiden University Medical Center; Leiden, the Netherlands
Transplant Center, Leiden University Medical Center; Leiden, the Netherlands
Department of Surgery, Leiden University Medical Center
Leiden, the Netherlands
Transplant Center, Leiden University Medical Center
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Leiden University Medical Center · Amsterdam University Medical Centers · University of Amsterdam
Affiliation as printed
Department of Pathology, Amsterdam UMC, University of Amsterdam; Amsterdam, the Netherlands
Department of Pathology, Leiden University Medical Center; Leiden, the Netherlands
Amsterdam, the Netherlands
Department of Pathology, Amsterdam UMC, University of Amsterdam
Department of Pathology, Leiden University Medical Center
Leiden, the Netherlands
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Leiden University Medical Center
Affiliation as printed
Department of Surgery, Leiden University Medical Center; Leiden, the Netherlands
Transplant Center, Leiden University Medical Center; Leiden, the Netherlands
Department of Surgery, Leiden University Medical Center
Leiden, the Netherlands
Transplant Center, Leiden University Medical Center
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Cees van Kooten Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine
Leiden University · Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center; Leiden, the Netherlands
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center
Leiden, the Netherlands
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Bram Heijs Aachen Center of Proteomics and Metabolomics Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Center of Proteomics and Metabolomics, Leiden University Medical Center; Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center; Leiden, the Netherlands
Center of Proteomics and Metabolomics, Leiden University Medical Center
Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center
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Gangqi Wang corresponding Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University · Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center; Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center; Leiden, the Netherlands
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center
Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center
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Marten A. Engelse corresponding Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University · Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center; Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center; Leiden, the Netherlands
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center
Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center
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Ton J. Rabelink corresponding Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University · Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center; Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center; Leiden, the Netherlands
Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center
Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center
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