University Hospital Zurich View Institution's Website 21 articles published in JoVE Neuroscience Ex Vivo OCT-Based Multimodal Imaging of Human Donor Eyes for Research into Age-Related Macular Degeneration Jeffrey D. Messinger1, Max Brinkmann2, James A. Kimble1, Andreas Berlin1, K. Bailey Freund3,4, Gregory H. Grossman5, Thomas Ach*6, Christine A. Curcio*1 1Department of Ophthalmology and Visual Sciences, University of Alabama at Birmingham Heersink School of Medicine, 2Department of Ophthalmology, University Hospital of Zurich, 3Vitreous Retina Macula Consultants of New York, 4Department of Ophthalmology, New York University Grossman School of Medicine, 5Advancing Sight Network, 6Department of Ophthalmology, University Hospital Bonn Laboratory assays can leverage prognostic value from the longitudinal optical coherence tomography (OCT)-based multimodal imaging of age-related macular degeneration (AMD). Human donor eyes with and without AMD are imaged using OCT, color, near-infrared reflectance scanning laser ophthalmoscopy, and autofluorescence at two excitation wavelengths prior to tissue sectioning. Bioengineering Simple Establishment of a Vascularized Osteogenic Bone Marrow Niche Using Pre-Cast Poly(ethylene Glycol) (PEG) Hydrogels in an Imaging Microplate Lisa A. Krattiger1, Maria Mitsi2, Benjamin R. Simona2, Martin Ehrbar1 1Department of Obstetrics, University Hospital Zurich, University of Zurich, 2Ectica Technologies AG An in vitro model of the bone marrow vascular niche is established by seeding mesenchymal and endothelial cells onto pre-cast 3D PEG hydrogels. The endothelial networks, ECM components, and ALP activity of the niches vary depending on the growth factor used. The platform can be used for advanced cancer models. Medicine Surgical Approach, Challenges, and Resolutions for Uterus Transplantation in Rats Keyue Sun1, Daniela Bochicchio1, Pierre-Alain Clavien1, Philipp Dutkowski1, Bostjan Humar1 1Swiss HPB Laboratory, Department of Visceral & Transplant Surgery, University Hospital Zürich The present protocol describes all the essential steps for successful uterine transplantation (UTx) in rats. The rat model has proven suitable to promote the clinical implementation of UTx; however, rat UTx is a highly complex procedure requiring careful instructions. Medicine Isolation of Regenerating Hepatocytes after Partial Hepatectomy in Mice Eva Breuer1, Bostjan Humar1 1Swiss HPB Laboratory, Department of Visceral & Transplant Surgery, University Hospital Zurich (USZ) Lipid-laden hepatocytes are inherent to liver regeneration but are usually lost upon density-gradient centrifugation. Here, we present an optimized cell isolation protocol that retains steatotic hepatocytes, yielding representative populations of regenerating hepatocytes after partial hepatectomy in mice. Medicine A Reproducible Intensive Care Unit-Oriented Endotoxin Model in Rats Jan Heil1, Martin Schläpfer1,2 1Institute of Physiology, University of Zurich, 2Institute of Anesthesiology, University Hospital of Zurich Here, we present a reproducible intensive care unit-oriented endotoxin model in rats. Medicine Clinical Practice Protocol of Creative Music Therapy for Preterm Infants and Their Parents in the Neonatal Intensive Care Unit Friederike B. Haslbeck1, Dirk Bassler1 1Department of Neonatology, University Hospital Zurich, University of Zurich Creative music therapy for preterm infants and their parents has emerged as a promising family-integrated early intervention. We present a detailed protocol on how to use vocal interaction, humming, or singing to empower preterm infants and their families. Cancer Research Desthiobiotin-Streptavidin-Affinity Mediated Purification of RNA-Interacting Proteins in Mesothelioma Cells Jelena Kresoja-Rakic1, Emanuela Felley-Bosco1 1Laboratory of Molecular Oncology, Division of Thoracic Surgery, University Hospital Zurich Desthiobiotin labeling of a synthetic 25-nucleotide RNA oligo, which contains an adenine-rich element (ARE) motif, allows specific binding of cytosolic ARE-binding protein. Developmental Biology Reprogramming Primary Amniotic Fluid and Membrane Cells to Pluripotency in Xeno-free Conditions Jaroslav Slamecka1, Javier Laurini2, Troy Shirley1, Simon Philipp Hoerstrup3, Benedikt Weber3,4,5, Laurie Owen1, Steven McClellan1 1Mitchell Cancer Institute, University of South Alabama, 2College of Medicine, University of South Alabama, 3Institute for Regenerative Medicine, University of Zurich, 4Department of Dermatology, University Hospital Zurich, 5Center for Applied Biotechnology and Molecular Medicine (CABMM), University of Zurich - Irchel Campus This protocol describes the reprogramming of primary amniotic fluid and membrane mesenchymal stem cells into induced pluripotent stem cells using a non-integrating episomal approach in fully chemically defined conditions. Procedures of extraction, culture, reprogramming, and characterization of the resulting induced pluripotent stem cells by stringent methods are detailed. Medicine Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata Simon Zimmermann1, Richard M. Fakin1, Thomas Giesen1, Pietro Giovanoli1, Maurizio Calcagni1 1Division of Plastic and Hand Surgery, University Hospital Zurich The purpose of the study is to illustrate the technical procedure of stromal vascular fraction (SVF)-enriched fat grafting as a novel treatment of symptomatic end-neuromata. This technique provides advantages of both the mechanical barrier and the biological action at the cellular level by the processed and concentrated SVF. Medicine Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure Erik Schadde1,2,3, Martin Hertl2, Stefan Breitenstein3, Beatrice Beck-Schimmer1,4, Martin Schläpfer1,4 1Institute of Physiology - Center for Integrative Human Physiology, University of Zurich, 2Department of Surgery, Rush University Medical Center, 3Department of Surgery, Cantonal Hospital Winterthur, 4Institute of Anesthesiology, University and University Hospital Zurich Inducing rapid liver hypertrophy using Associating Liver Partition and Portal vein ligation for a Staged hepatectomy (ALPPS) has been proposed for resection of borderline resectable liver tumors. This model may elucidate mechanisms involved in rapid hypertrophy and allows testing of drugs that promote or block the acceleration of regeneration. Bioengineering Isolation of Murine Adipose Tissue-derived Microvascular Fragments as Vascularization Units for Tissue Engineering Florian S. Frueh1,2, Thomas Später1, Claudia Scheuer1, Michael D. Menger1, Matthias W. Laschke1 1Institute for Clinical and Experimental Surgery, Saarland University, 2Division of Plastic Surgery and Hand Surgery, University Hospital Zurich, University of Zurich We present a protocol to isolate adipose tissue-derived microvascular fragments that represent promising vascularization units. They can be rapidly isolated, do not require in vitro processing and, thus, may be used for one-step prevascularization in different fields of tissue engineering. Behavior Investigating Motor Skill Learning Processes with a Robotic Manipulandum Susan Leemburg1, Maiko Iijima1, Olivier Lambercy2, Lauriane Nallet-Khosrofian1, Roger Gassert2, Andreas Luft1 1Division of Vascular Neurology and Rehabilitation, Department of Neurology, University Hospital Zurich, 2Rehabilitation Engineering Laboratory, Department of Health Sciences and Technology, ETH Zurich A paradigm is presented for training and analysis of an automated skilled reaching task in rats. Analysis of pulling attempts reveals distinct subprocesses of motor learning. Developmental Biology An Enzyme- and Serum-free Neural Stem Cell Culture Model for EMT Investigation Suited for Drug Discovery Martin H. M. Sailer1, Durga Sarvepalli2, Catherine Brégère3, Urs Fisch3, Marin Guentchev4, Michael Weller6, Raphael Guzman3, Bernhard Bettler1, Arkasubhra Ghosh*2, Gregor Hutter*5 1Dept. of Biomedicine, Pharmacenter, University of Basel, 2Molecular Signalling and Gene Therapy, Narayana Nethralaya Foundation, Narayana Health City, 3Brain Ischemia and Regeneration, Department of Biomedicine, University Hospital Basel, 4Department of Neurosurgery, Klinikum Idar-Oberstein, 5Department of Neurosurgery and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, 6Department of Neurology, Laboratory of Molecular Neuro Oncology, University Hospital of Zurich Epithelial to mesenchymal transition (EMT) allows cancers to become invasive. To investigate EMT, a neural stem cell (NSC)-based in vitro model devoid of serum and enzymes is described. This standardized system allows quantitative and qualitative assessment of cell migration, gene and protein expression. The model is suited for drug discovery. Medicine Robotic Ablation of Atrial Fibrillation Alexander Wutzler1, Thomas Wolber2, Wilhelm Haverkamp1, Leif-Hendrik Boldt1 1Department of Cardiology, Charité — Universitätsmedizin Berlin, Campus Virchow, 2Department of Cardiology, University Hospital Zurich Pulmonary vein isolation (PVI) with an ablation catheter is a curative treatment for atrial fibrillation (AF). Robotic catheter systems aim to improve catheter steerability. Here, a procedure with a new robotic catheter system is presented. The goal of the procedure is electrical block between pulmonary vein and left atrium. Medicine Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia Yves Harder1, Daniel Schmauss1, Reto Wettstein2, José T. Egaña1, Fabian Weiss1, Andrea Weinzierl1, Anna Schuldt1, Hans-Günther Machens1, Michael D. Menger3, Farid Rezaeian4 1Department of Plastic Surgery and Hand Surgery, Klinikum rechts der Isar, Technische Universität München, 2Department of Plastic, Reconstructive, Aesthetic and Hand Surgery, University Hospital of Basel, 3Institute for Clinical and Experimental Surgery, University of Saarland, 4Division of Plastic and Hand Surgery, University Hospital Zurich The window of the murine dorsal skinfold chamber presented visualizes a zone of acute persistent ischemia of a musculocutaneous flap. Intravital epi-fluorescence microscopy permits for direct and repetitive assessment of the microvasculature and quantification of hemodynamics. Morphologic and hemodynamic results can further be correlated with histological and molecular analyses. Bioengineering A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo Thilo L. Schenck1, Myra N. Chávez1, Alexandru P. Condurache2, Ursula Hopfner1, Farid Rezaeian3, Hans-Günther Machens1, José T. Egaña1,4 1Department of Plastic Surgery and Hand Surgery, University Hospital rechts der Isar, Technische Universität München, 2Institute for Signal Processing, University of Lübeck, 3Department of Plastic Surgery and Hand Surgery, University Hospital Zürich, 4FONDAP Center for Genome Regulation, Facultad de Ciencias, Universidad de Chile Vascularization is key to approaches in successful tissue engineering. Therefore, reliable technologies are required to evaluate the development of vascular networks in tissue-constructs. Here we present a simple and cost-effective method to visualize and quantify vascularization in vivo. Bioengineering Preparation of Light-responsive Membranes by a Combined Surface Grafting and Postmodification Process Katrin Schöller1, Lukas Baumann1, Dirk Hegemann2, Damien De Courten3, Martin Wolf3, René M. Rossi1, Lukas J. Scherer1 1Laboratory for Protection and Physiology, Empa, Swiss Federal Laboratories for Materials Science and Technology, 2Laboratory of Advanced Fibers, Empa, Swiss Federal Laboratories for Materials Science and Technology, 3Division of Neonatology, University Hospital Zurich A plasma-induced polymerization procedure is described for the surface-initiated polymerization on polymer membranes. Further postmodification of the grafted polymer with photochromic substances is presented with a protocol of conducting permeability measurements of light-responsive membranes. Immunology and Infection Intralymphatic Immunotherapy and Vaccination in Mice Pål Johansen1, Thomas M. Kündig1 1Department of Dermatology, University Hospital Zurich Prophylactic and therapeutic vaccination often fails to stimulate strong immune responses due to week drainage of the vaccine to lymph nodes and consequently poor involvement of immune cells. By direct injection of vaccine to lymph nodes, so-called intralymphatic injection, vaccine efficacy can be strongly improved and vaccine doses can be reduced. Bioengineering Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model Stefanie Grafmüller1,2,3, Pius Manser2, Harald F. Krug2, Peter Wick2, Ursula von Mandach1 1Department of Obstetrics, Perinatal Pharmacology, University Hospital Zurich, 2Laboratory for Materials - Biology Interactions, EMPA Swiss Federal Laboratories for Materials Testing and Research, 3Graduate School for Cellular and Biomedical Sciences, University of Bern The ex vivo dual recirculating human placental perfusion model can be used to investigate the transfer of xenobiotics and nanoparticles across the human placenta. In this video protocol we describe the equipment and techniques required for a successful execution of a placenta perfusion. Medicine Roux-en-Y Gastric Bypass Operation in Rats Marco Bueter1,2, Kathrin Abegg2,3, Florian Seyfried4, Thomas A. Lutz2,3, Carel W. le Roux4 1Department of Surgery, University Hospital Zürich, 2Zürich Centre for Integrative Human Physiology, University of Zürich, 3Institute of Veterinary Physiology, Vetsuisse Faculty, University of Zürich, 4Imperial Weight Centre, Department of Investigative Medicine, Imperial College London Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool. Medicine Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice Nikola Cesarovic1, Paulin Jirkof2, Andreas Rettich2, Margarete Arras1 1Division of Surgical Research, University Hospital Zurich, 2Institute of Laboratory Animal Science, University of Zurich A surgical technique for implantation of commercially available telemetry transmitters used for continuous measurement of biopotential (one-lead ECG), heart rate, core body temperature and locomotor activity in freely moving mice is shown. Recommendations and protocols for post-operative care and pain relief, improving recovery, well being and survival rate are also presented.