University Rostock Medical Center 9 articles published in JoVE Cancer Research Creation and Maintenance of a Living Biobank - How We Do It Florian Bürtin1, Stephanie Matschos2, Friedrich Prall3, Christina S. Mullins2, Mathias Krohn2, Michael Linnebacher2 1Department of General, Visceral, Vascular and Transplantation Surgery, University Medical Center Rostock, University of Rostock, 2Molecular Oncology and Immunotherapy, Department of General, Visceral, Vascular and Transplantation Surgery, University Medical Center Rostock, 3Institute of Pathology, University Medical Center Rostock In the following work, we describe the consecutive steps necessary for the establishment of a large biobank of colorectal and pancreatic cancer. Medicine Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion Christoph R. Behem1, Till Friedheim1, Sabine H. Wipper2, Hans O. Pinnschmidt3, Michael F. Graessler1, Catharina Gaeth4, Hannes Holthusen1, Adina Rapp5, Timo Suntrop1, Josephina Haunschild6, Christian D. Etz6, Constantin J. C. Trepte1 1Department of Anesthesiology, Center of Anesthesiology and Intensive Care Medicine, University Medical Center Hamburg-Eppendorf, 2University Department for Vascular Surgery and Department of Operative Medicine, Medical University of Innsbruck, 3Department of Medical Biometry and Epidemiology, University Medical Center Hamburg-Eppendorf, 4Department of Vascular Medicine, University Heart and Vascular Center Hamburg (UHZ), 5Department of Cardiology, Rostock University Medical Center, 6University Department for Cardiac Surgery, Heart Center Leipzig Spinal cord microcirculation plays a pivotal role in spinal cord injury. Most methods do not allow real-time assessment of spinal cord microcirculation, which is essential for the development of microcirculation-targeted therapies. Here, we propose a protocol using Laser-Doppler-Flow Needle probes in a large animal model of ischemia/reperfusion. Biology Analyzing the α-Actinin Network in Human iPSC-Derived Cardiomyocytes Using Single Molecule Localization Microscopy Lisa Johann1,2, Oleksandra Chabanovska1,2, Cajetan Immanuel Lang3, Robert David1,2, Heiko Lemcke1,2 1Department of Cardiac Surgery, Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Rostock University Medical Center, 2Faculty of Interdisciplinary Research, Department Life, Light & Matter, University Rostock, 3Department of Cardiology, Rostock University Medical Center The formation of a proper sarcomere network is important for the maturation of iPSC-derived cardiomyocytes. We present a super resolution-based approach that allows for the quantitative evaluation of the structural maturation of stem cell derived cardiomyocytes, to improve culture conditions promoting cardiac development. Medicine Intrathecal Application of a Fluorescent Dye for the Identification of Cerebrospinal Fluid Leaks in Cochlear Malformation Nora M. Weiss1, Ingo Andus2, Armin Schneider3, Sönke Langner4, Stefanie Schröder1, Sebastian P. Schraven1, Robert Mlynski1 1Department of Otorhinolaryngology, Head and Neck Surgery, University Medical Center Rostock, 2Rostock University, 3ARRI Medical GmbH, 4Department of Radiology and Institute of Diagnostic and Interventional Radiology, University Medical Center Rostock Intrathecally applied fluorescein is used to achieve intraoperative visualization of CSF leaks. This protocol describes a lumbar puncture, the application of 5% fluorescein, and intraoperative visualization using a fully digital microscope. Medicine Continuous Blood Sampling in Small Animal Positron Emission Tomography/Computed Tomography Enables the Measurement of the Arterial Input Function Teresa Mann1, Jens Kurth2, Anne Möller3, Joanna Förster3, Brigitte Vollmar4, Bernd J. Krause2, Andreas Wree1, Jan Stenzel*3, Tobias Lindner*3 1Institute of Anatomy, Rostock University Medical Center, 2Department of Nuclear Medicine, Rostock University Medical Center, 3Core Facility Multimodal Small Animal Imaging, Rostock University Medical Center, 4Rudolf-Zenker-Institute for Experimental Surgery, Rostock University Medical Center Here a protocol for continuous blood sampling during PET/CT imaging of rats to measure the arterial input function (AIF) is described. The catheterization, the calibration and setup of the system and the data analysis of the blood radioactivity are demonstrated. The generated data provide input parameters for subsequent bio-kinetic modeling. Genetics Isolation, Characterization and MicroRNA-based Genetic Modification of Human Dental Follicle Stem Cells Paula Müller*1,2, Katharina Ekat*3, Anne Brosemann3, Anne Köntges3, Robert David1,2, Hermann Lang3 1Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, 2Department Life, Light and Matter of the Interdisciplinary Faculty at Rostock University, 3Department of Operative Dentistry and Periodontology, Rostock University Medical Center This protocol describes the transient genetic engineering of dental stem cells extracted from the human dental follicle. The applied non-viral modification strategy may become a basis for the improvement of therapeutic stem cell products. Bioengineering Protocol for MicroRNA Transfer into Adult Bone Marrow-derived Hematopoietic Stem Cells to Enable Cell Engineering Combined with Magnetic Targeting Frauke Hausburg*1,2, Paula Müller*1,2, Natalia Voronina*1, Gustav Steinhoff1,2, Robert David1,2 1Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, 2Department Life, Light and Matter of the Interdisciplinary Faculty, Rostock University This protocol illustrates a safe and efficient procedure to modify CD133+ hematopoietic stem cells. The presented non-viral, magnetic polyplex-based approach may provide a basis for the optimization of therapeutic stem cell effects as well as for monitoring the administered cell product via magnetic resonance imaging. Medicine In Vitro Enzyme Measurement to Test Pharmacological Chaperone Responsiveness in Fabry and Pompe Disease Jan Lukas1, Anne-Marie Knospe1, Susanne Seemann1, Valentina Citro2, Maria V. Cubellis2, Arndt Rolfs1,3 1Albrecht-Kossel-Institute, University Rostock Medical Center, 2Department of Biology, University Federico II, 3Centogene AG There is a demand to make pre-clinical testing for a novel class of "orphan" drugs called pharmacological chaperones reproducible, fast, and efficient. We developed a simple, highly standardized, and versatile cell culture-based assay to screen for eligible patients as well as novel pharmacological chaperone drugs. Medicine Intravital Microscopy and Thrombus Induction in the Earlobe of a Hairless Mouse Daniel Strüder*1, Eberhard Grambow*2, Ernst Klar2, Robert Mlynski1, Brigitte Vollmar3 1Department of Otorhinolaryngology, Head and Neck Surgery "Otto Koerner", Rostock University Medical Center, 2Department of General, Thoracic, Vascular and Transplantation Surgery, Rostock University Medical Center, 3Institute for Experimental Surgery, Rostock University Medical Center The ear model of the hairless SKH1-Hrhr mouse enables intravital fluorescence microscopy of microcirculation and phototoxic thrombus induction without prior surgical preparation in the examined microvascular bed. Therefore, the ear of the hairless mouse is an excellent in vivo model to study the complex interactions during microvascular thrombus formation, thrombus evolution, and thrombolysis.