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University Hospital RWTH Aachen

12 ARTICLES PUBLISHED IN JoVE

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Medicine

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats
Karina Schleimer 1, Jochen Grommes 1, Andreas Greiner 1, Houman Jalaie 1, Johannes Kalder 1, Stephan Langer 1, Thomas A. Koeppel 1, Michael Jacobs 1, Maria Kokozidou 1
1European Vascular Center Aachen-Maastricht, Department of Vascular Surgery, University Hospital RWTH Aachen

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

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Medicine

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis
Sakine Simsekyilmaz 1, Fabian Schreiber 2, Stefan Weinandy 3, Felix Gremse 4, Tolga Taha Sönmez 5, Elisa A. Liehn 1
1Institute for Molecular Cardiovascular Research, RWTH Aachen University, 2Institute for Textile Technology and Mechanical Engineering, RWTH Aachen University, 3Institute for Applied Medical Engineering, Helmholtz-Institute of RWTH Aachen University, 4Department of Experimental Molecular Imaging, RWTH Aachen University, 5Department of Oral and Maxillofacila Surgery, RWTH Aachen University

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

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Medicine

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure
Karina Schleimer 1, Johannes Kalder 1, Jochen Grommes 1, Houman Jalaie 1, Samir Tawadros 1, Andreas Greiner 1, Michael Jacobs 1, Maria Kokozidou 1
1European Vascular Center Aachen-Maastricht, Department of Vascular Surgery, University Hospital RWTH Aachen

Auxiliary liver transplantation provides a temporary support in acute hepatic failure, until regeneration of the failing liver. The heterotopic auxiliary liver transplantation (HALT) with portal vein arterialization (PVA) renders sufficient liver function. We developed an analogous technique in the rat, to examine the influence of the portal vein arterialization on the morphology and function of the graft.

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Medicine

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice
Adelina Curaj 1, Sakine Simsekyilmaz 1, Mareike Staudt 1, Elisa Liehn 1
1Institute for Molecular Cardiovascular Research, RWTH Aachen University, Germany

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

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Medicine

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video
Zuzanna Rowinska 1,2, Simone Gorressen 3, Marc W. Merx 4, Thomas A. Koeppel 5, Alma Zernecke 6, Elisa A. Liehn 2
1Department of Vascular Surgery and Interdisciplinary Vein Center, St. Josef-Hospital, Ruhr- University Bochum, 2Institute of Molecular Cardiovascular Research, University Hospital, RWTH Aachen University Department of Medicine, 3Institute of Pharmacology and Clinical Pharmacology, Düsseldorf University Hospital, 4Department of Cardiology, Vascular Medicine and Intensive Care Medicine, Hospital Robert Koch Gehrden, 5Division of Vascular Surgery, Asklepios Klinik St. Georg, 6Institute of Experimental Biomedicine, Würzburg University

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

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Medicine

Optical Clearing and Imaging of Immunolabeled Kidney Tissue
Turgay Saritas 1, Victor G. Puelles 1,2,3, Xiao-Tong Su 4, David H. Ellison 4,5,6, Rafael Kramann 1,7
1Division of Nephrology and Clinical Immunology, University Hospital RWTH Aachen, 2III. Department of Medicine, University Medical Center, Hamburg-Eppendorf, 3Department of Nephrology, Monash Health, 4Division of Nephrology and Hypertension, Oregon Health and Science University, 5Renal Section, Veterans Affairs Portland Health Care System, 6Fondation LeDucq Transatlantic Networks of Excellence, 7Department of Internal Medicine, Nephrology and Transplantation, Erasmus Medical Center

The combination of antibody labeling, optical clearing, and advanced light microscopy allows three-dimensional analysis of complete structures or organs. Described here is a simple method to combine immunolabeling of thick kidney slices, optical clearing with ethyl cinnamate, and confocal imaging that enables visualization and quantification of three-dimensional kidney structures.

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Medicine

Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access
Anne Cornelissen *1, Roberta Florescu *1, Nicole Schaaps 1, Mamdouh Afify 1, Sakine Simsekyilmaz 1, Elisa Liehn *1,2,3, Felix Vogt *1
1Division of Cardiology, Angiology, and Critical Care, University Hospital RWTH Aachen, 2Department for Operative Intensive Medicine, University Hospital RWTH Aachen, 3Institute for Pathology "Victor Babes"

This protocol describes the implantation of human coronary stents into the abdominal aorta of rats with an apoE-/- background using a trans-femoral access. Compared with other animal models, murine models carry the advantages of high throughput, reproducibility, ease of handling and housing, and a broad availability of molecular markers.

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Medicine

Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry
Wen-Jia Liu *1,2, Lisa Ernst *2, Benedict Doorschodt 2, Jan Bednarsch 1, Felix Becker 3, Richi Nakatake 2, Yuki Masano 2, Ulf Peter Neumann 1,4, Sven Arke Lang 1, Peter Boor 5, Isabella Lurje 6, Georg Lurje 1,6, René Tolba 2, Zoltan Czigany 1,2
1Department of Surgery and Transplantation, Faculty of Medicine, University Hospital RWTH Aachen, 2Institute for Laboratory Animal Science, Faculty of Medicine, University Hospital RWTH Aachen, 3Department of General-, Visceral-, and Transplantation Surgery, University Hospital Muenster, 4Department of Surgery, Maastricht University Medical Centre (MUMC), 5Institute of Pathology, Faculty of Medicine, University Hospital RWTH Aachen, 6Department of Surgery, Campus Charité Mitte | Campus Virchow-Klinikum, Charité-Universitätsmedizin

Large animal models play an essential role in preclinical transplantation research. Due to its similarities to the clinical setup, the porcine model of orthotopic kidney auto-transplantation described in this article provides an excellent in vivo setting for the testing of organ preservation techniques and therapeutic interventions.

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Medicine

Electroporation-Based Genetic Modification of Primary Human Pigment Epithelial Cells Using the Sleeping Beauty Transposon System
Sandra Johnen 1, Nina Harmening 2,3, Corinne Marie 4,5, Daniel Scherman 4, Zsuzsanna Izsvák 6, Zoltán Ivics 7, Peter Walter 1, Gabriele Thumann 2,3
1Department of Ophthalmology, University Hospital RWTH Aachen, 2Experimental Ophthalmology, University of Geneva, 3Department of Ophthalmology, University Hospitals of Geneva, 4Université de Paris, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, 5Chimie ParisTech, PSL Research University, 6Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 7Division of Medical Biotechnology, Paul-Ehrlich-Institute

We have developed a protocol to transfect primary human pigment epithelial cells by electroporation with the gene encoding pigment epithelium-derived factor (PEDF) using the Sleeping Beauty (SB) transposon system. Successful transfection was demonstrated by quantitative polymerase chain reaction (qPCR), immunoblotting, and enzyme-linked immunosorbent assay (ELISA).

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JoVE Core

Cardiac Response to β-Adrenergic Stimulation Determined by Pressure-Volume Loop Analysis
Rebekka Medert *1,2, Lucas Bacmeister *1,2,3, Sebastian Segin *1,2,4, Marc Freichel 1,2, Juan E. Camacho Londoño 1,2
1Pharmakologisches Institut, Ruprecht-Karls-Universität Heidelberg, 2DZHK (German Centre for Cardiovascular Research), partner site Heidelberg/Mannheim, 3Department of Internal Medicine III, University of Heidelberg, 4Department of Anesthesiology, University Hospital RWTH Aachen

Here we describe a cardiac pressure-volume loop analysis under increasing doses of intravenously infused isoproterenol to determine the intrinsic cardiac function and the β-adrenergic reserve in mice. We use a modified open-chest approach for the pressure-volume loop measurements, in which we include ventilation with positive end-expiratory pressure.

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Biology

Isolation, Culture, and Genetic Engineering of Mammalian Primary Pigment Epithelial Cells for Non-Viral Gene Therapy
Thais Bascuas *1,2, Martina Kropp *1,2, Nina Harmening 1,2, Brittany M. Wong 1, Sandra Johnen 3, Zsuzsanna Izsvák 4, Gabriele Thumann 1,2
1Experimental Ophthalmology, University of Geneva, 2Department of Ophthalmology, University Hospitals of Geneva, 3Department of Ophthalmology, University Hospital RWTH Aachen, 4Max Delbrück Center for Molecular Medicine

Here, a protocol to isolate and transfect primary iris and retinal pigment epithelial cells from various mammals (mice, rat, rabbit, pig, and bovine) is presented. The method is ideally suited to study ocular gene therapy approaches in various set-ups for ex vivo analyses and in vivo studies transferable to humans.

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Cancer Research

Radiosynthesis, Quality Control, and Small Animal Positron Emission Tomography Imaging of 68Ga-labelled Nano Molecules
Betül Altunay 1, Matthias Bauwens 1,2,3, Laura Schäfer 1, Amelie Heesch 1, Agnieszka Morgenroth 1, Felix M. Mottaghy 1,2,4, Alexandru Florea 1,2,4
1Department of Nuclear Medicine, University Hospital RWTH Aachen, 2Department of Radiology and Nuclear Medicine, Maastricht University Medical Center, 3School of Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University, 4School for Cardiovascular Diseases (CARIM), Maastricht University

This paper describes the radiosynthesis, formulation, quality control of a new radiolabeled probe (i.e., 68Ga-labeled nanobody NM-02), and its use for small animal PET/CT imaging in a xenograft model.

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