University Hospital RWTH Aachen 6 articles published in JoVE Medicine 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 Freichel1,2, Juan E. Camacho Londoño1,2 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. 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 Harmening1,2, Brittany M. Wong1, Sandra Johnen3, Zsuzsanna Izsvák4, Gabriele Thumann1,2 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. Medicine Electroporation-Based Genetic Modification of Primary Human Pigment Epithelial Cells Using the Sleeping Beauty Transposon System Sandra Johnen1, Nina Harmening2,3, Corinne Marie4,5, Daniel Scherman4, Zsuzsanna Izsvák6, Zoltán Ivics7, Peter Walter1, Gabriele Thumann2,3 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). Medicine Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access Anne Cornelissen*1, Roberta Florescu*1, Nicole Schaaps1, Mamdouh Afify1, Sakine Simsekyilmaz1, 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. 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 Doorschodt2, Jan Bednarsch1, Felix Becker3, Richi Nakatake2, Yuki Masano2, Ulf Peter Neumann1,4, Sven Arke Lang1, Peter Boor5, Isabella Lurje6, Georg Lurje1,6, René Tolba2, Zoltan Czigany1,2 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. Medicine Optical Clearing and Imaging of Immunolabeled Kidney Tissue Turgay Saritas1, Victor G. Puelles1,2,3, Xiao-Tong Su4, David H. Ellison4,5,6, Rafael Kramann1,7 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.
