University Hospital Erlangen 8 articles published in JoVE Biology Generation of Patient-Derived Podocytes from Skin Biopsies Victoria Rose1, Janina Müller-Deile1,2 1Department of Nephrology and Hypertension, University Hospital Erlangen, Friedrich-Alexander University (FAU) Erlangen-Nürnberg, 2Research Center on Rare Kidney Diseases (RECORD), University Hospital Erlangen This manuscript describes a two-step protocol to generate patient-specific podocytes from dermal fibroblasts via episomal reprogramming into human-induced pluripotent stem cells (hiPSCs) and subsequent differentiation into podocytes. Immunology and Infection An Intravital Microscopy-Based Approach to Assess Intestinal Permeability and Epithelial Cell Shedding Performance Luz DC. Martínez-Sánchez*1, Rashmita Pradhan*1, Phuong A. Ngo1, Lena Erkert1, Lukas S. Becker1, Alastair J. Watson2, Imke Atreya*1, Markus F. Neurath*1, Rocío López-Posadas*1 1Medical Clinic 1, University of Erlangen-Nuremberg, University Hospital of Erlangen, 2Norwich Medical School, University of East Anglia, Norwich Research Park Taking advantage of intravital microscopy, the method presented here enables real-time visualization of intestinal epithelial cell shedding in living animals. Therefore, topically stained intestinal mucosa (acriflavine and rhodamineB-dextran) of anesthetized mice is imaged up to single-cell resolution using confocal microscopy. Medicine Cooling or Warming the Esophagus to Reduce Esophageal Injury During Left Atrial Ablation in the Treatment of Atrial Fibrillation Jason Zagrodzky1, Mark M. Gallagher2, Lisa W. M. Leung2, Tiffany Sharkoski3, Pasquale Santangeli4, Cory Tschabrunn4, Jose M. Guerra5, Bieito Campos5, John MacGregor6, Jamal Hayat2, Brad Clark7, Alex Mazur8, Marcel Feher9, Martin Arnold9, Mark Metzl10, Jose Nazari10, Erik Kulstad11 1 The goal of this protocol is to describe the use of esophageal temperature modulation to counteract esophageal thermal injury from left atrial ablation for the treatment of atrial fibrillation. Medicine Advanced Imaging of Lung Homing Human Lymphocytes in an Experimental In Vivo Model of Allergic Inflammation Based on Light-sheet Microscopy Anja Schulz-Kuhnt1, Sebastian Zundler1, Anika Grüneboom2, Clemens Neufert1, Stefan Wirtz1, Markus F. Neurath1, Imke Atreya1 1Department of Medicine 1, University Hospital of Erlangen, 2Department of Medicine 3, University Hospital of Erlangen The here introduced protocol allows characterization of the lung homing capacity of primary human lymphocytes under in vivo inflammatory conditions. Pulmonary infiltration of adoptively transferred human immune cells in a mouse model of allergic inflammation can be imaged and quantified by light-sheet fluorescence microscopy of chemically cleared lung tissue. Immunology and Infection Induction of Intestinal Graft-versus-host Disease and Its Mini-endoscopic Assessment in Live Mice Vera Buchele1,3, Maike Büttner-Herold2, Tina Vogler1,3, Markus F. Neurath1,3, Kai Hildner1,3 1Department of Internal Medicine 1, University Erlangen-Nürnberg, University Hospital Erlangen, 2Institute of Pathology, Department of Nephropathology, University Hospital Erlangen, 3Deutsches Zentrum Immuntherapie (DZI), University Hospital Erlangen Here, we present a protocol that describes allogeneic hematopoietic stem cell transplantation and allows repetitive mini-endoscopic evaluations of the distal colon in situ for the presence, characteristics, and severity of colonic inflammation within live mice suffering from intestinal graft-versus-host disease. Medicine The Influence of Liver Resection on Intrahepatic Tumor Growth Hannes H. Brandt1, Valérie Nißler2, Roland S. Croner3 1Friedrich-Alexander-University Erlangen-Nürnberg (FAU), 2Department of Surgery, University Hospital Regensburg, 3Department of Surgery, University Hospital Erlangen A high incidence of tumor recurrence after resection of liver metastases remains an unsolved problem. The illustrated mouse model may be useful to investigate the reasons for such recurrences. It combines a liver resection model with intrahepatic tumor cell injection for the first time. Medicine Surgical Fixation of Sternal Fractures: Preoperative Planning and a Safe Surgical Technique Using Locked Titanium Plates and Depth Limited Drilling Stefan Schulz-Drost1, Pascal Oppel1, Sina Grupp1, Sonja Schmitt1, Roman Th. Carbon2, Andreas Mauerer3, Friedrich F. Hennig1, Thomas Buder4 1Orthopedic and Trauma Surgery, University Hospital Erlangen, 2Pediatric Surgery, University Hospital Erlangen, 3Orthopedic and Trauma Surgery, St.-Theresien Hospital, 4Institute of Anatomy I, University Erlangen-Nuremberg Here we present a method to stabilize sternal fractures by using locked titanium plates in a low profile design. Performing subperiosteal dissection along the sternum while reducing the fracture, using depth limited drilling, and fixing the plates provides a safe surgical way. Immunology and Infection Isolation of Myeloid Dendritic Cells and Epithelial Cells from Human Thymus Christina Stoeckle1,2, Ioanna A. Rota1, Eva Tolosa3, Christoph Haller4, Arthur Melms5, Eleni Adamopoulou1 1Department of General Neurology, Hertie Institute for Clinical Brain Research, 2Institute of Pharmacology, University of Bern, 3Department of Immunology, University Medical Center Hamburg-Eppendorf, 4Department of Thoracic and Cardiovascular Surgery, University Clinic Tuebingen, 5Department of Neurology, University Hospital Erlangen This protocol details a method to isolate antigen presenting cells from human thymus via different steps of enzymatic digestion of the tissue followed by density centrifugation of the single cell suspension and finally magnetic and/or FACS sorting of the cell populations of interest.
