Technical University of Munich (TUM) 13 articles published in JoVE Biology Visualizing Scar Development Using SCAD Assay - An Ex-situ Skin Scarring Assay Pushkar Ramesh1, Haifeng Ye1, Bikram Dasgupta1, Hans-Günther Machens2, Yuval Rinkevich1 1Institute of Regenerative Biology and Medicine, Helmholtz Zentrum München, 2School of Medicine, Klinikum Rechts der Isar, Department of Plastic and Hand Surgery, Technical University of Munich This protocol describes the generation of a skin-fascia explant termed "SCar like tissue in A Dish" or SCAD. This model allows unprecedented visualization of single fibroblasts during scar formation. Cancer Research In Vivo Immunogenicity Screening of Tumor-Derived Extracellular Vesicles by Flow Cytometry of Splenic T Cells Florian Stritzke1,2, Hendrik Poeck1,2,3,4, Simon Heidegger1,2 1Department of Medicine III, School of Medicine, Technical University of Munich, 2Center for Translational Cancer Research (TranslaTUM), School of Medicine, Technical University of Munich, 3Department of Internal Medicine III, University Hospital Regensburg, 4National Centre for Tumor Diseases WERA This manuscript describes how to assess in vivo immunogenicity of tumor cell-derived extracellular vesicles (EVs) using flow cytometry. EVs derived from tumors undergoing treatment-induced immunogenic cell death seem particularly relevant in tumor immunosurveillance. This protocol exemplifies the assessment of oxaliplatin-induced immunostimulatory tumor EVs but can be adapted to various settings. Neuroscience Preparing Adult Drosophila melanogaster for Whole Brain Imaging during Behavior and Stimuli Responses Alexandra Woller1, Paul Bandow1, Sophie Aimon1, Ilona C. Grunwald Kadow1 1Chair of Neuronal Control and Metabolism, School of Life Sciences, Technical University of Munich We present a method specifically tailored to image the whole brain of adult Drosophila during behavior and in response to stimuli. The head is positioned to allow optical access to the whole brain, while the fly can move its legs and the antennae, the tip of the proboscis, and the eyes can receive sensory stimuli. Medicine Development and Evaluation of 3D-Printed Cardiovascular Phantoms for Interventional Planning and Training Maximilian Grab1,2, Carina Hopfner3, Alena Gesenhues4, Fabian König1,2, Nikolaus A. Haas3, Christian Hagl1, Adrian Curta4, Nikolaus Thierfelder1 1Department of Cardiac Surgery, Ludwig Maximilian University Munich, 2Chair of Medical Materials and Implants, Technical University of Munich, 3Department Pediatric Cardiology and Pediatric Intensive Care, Ludwig Maximilian University Munich, 4Department of Radiology, Ludwig Maximilian University Munich Here we present development of a mock circulation setup for multimodal therapy evaluation, pre-interventional planning, and physician-training on cardiovascular anatomies. With the application of patient-specific tomographic scans, this setup is ideal for therapeutic approaches, training, and education in individualized medicine. Bioengineering 3D Imaging of Soft-Tissue Samples using an X-ray Specific Staining Method and Nanoscopic Computed Tomography Madleen Busse*1, Mark Müller*1, Melanie A. Kimm2, Simone Ferstl1, Sebastian Allner1, Klaus Achterhold1, Julia Herzen1, Franz Pfeiffer1,2 1Department of Physics and Munich School of BioEngineering, Technical University of Munich, 2Department of Diagnostic and Interventional Radiology, School of Medicine and Klinikum rechts der Isar, Technical University of Munich A protocol for 3D visualization of microscopic tissue structures by using an X-ray specific staining method designed for X-ray computed tomography is presented. Bioengineering In Vesiculo Synthesis of Peptide Membrane Precursors for Autonomous Vesicle Growth Kilian Vogele1, Thomas Frank1, Lukas Gasser1, Marisa A. Goetzfried1, Mathias W. Hackl2, Stephan A. Sieber2, Friedrich C. Simmel1,3, Tobias Pirzer1 1Physics of Synthetic Systems - E14, Physics-Department and ZNN, Technische Universität München, 2Departmant of Chemistry, Center for Integrated Protein Science (CIPSM), Technische Universität München, 3Nanosystems Initiative Munich Presented here are protocols for the creation of peptide-based small unilamellar vesicles capable of growth. To facilitate in vesiculo production of the membrane peptide, these vesicles are equipped with a transcription-translation system and the peptide-encoding plasmid. Bioengineering Functional Surface-immobilization of Genes Using Multistep Strand Displacement Lithography Günther Pardatscher*1, Matthaeus Schwarz-Schilling*1, Sandra Sagredo1, Friedrich C. Simmel1 1Physics Department, Technical University of Munich We describe a simple lithographic procedure for the immobilization of gene-length DNA molecules on a surface, which can be used to perform cell-free gene expression experiments on biochips. Biology Subtype-specific Optical Action Potential Recordings in Human Induced Pluripotent Stem Cell-derived Ventricular Cardiomyocytes Alexander Goedel*1,2, Dorota M. Zawada*1, Fangfang Zhang1, Zhifen Chen3, Alessandra Moretti1,2, Daniel Sinnecker1,2 1Medical Department I, University Hospital Klinikum rechts der Isar, Technical University of Munich, 2German Centre for Cardiovascular Research (DZHK), Munich Heart Alliance, 3Beth Israel Deaconess Medical Center, Harvard Medical School Here we present a method to optically image action potentials, specifically in ventricular-like induced pluripotent stem cell-derived cardiomyocytes. The method is based on the promoter-driven expression of a voltage-sensitive fluorescent protein. Cancer Research Therapy Testing in a Spheroid-based 3D Cell Culture Model for Head and Neck Squamous Cell Carcinoma Jan Hagemann*1, Christian Jacobi*2, Sabine Gstoettner3, Christian Welz4, Sabina Schwenk-Zieger3, Roland Stauber1, Sebastian Strieth1, Julian Kuenzel1, Philipp Baumeister3, Sven Becker1,3 1Department of Otorhinolaryngology, Johannes-Gutenberg University Medical Center, 2Department of Otorhinolaryngology, Technical University of Munich Medical Center, 3Department of Otorhinolaryngology, Ludwig-Maximilian-University Medical Center, 4Department of Otorhinolaryngology, University of Goettingen Medical Center We describe the evolution of a spheroid-based, three-dimensional in vitro model that enables us to test the current standard of experimental therapy regimens for head and neck squamous cell carcinoma on cell lines, aiming at evaluating therapy susceptibility and resistance on primary cells from human specimens in the future. Bioengineering Melt Electrospinning Writing of Three-dimensional Poly(ε-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications Felix M. Wunner1, Onur Bas1, Navid T. Saidy1, Paul D. Dalton2, Elena M. De-Juan Pardo1, Dietmar W. Hutmacher1,3,4 1ARC ITTC in Additive Biomanufacturing, Institute for Health and Biomedical Innovation (IHBI), Queensland University of Technology (QUT), 2Department for Functional Materials in Medicine and Dentistry and Bavarian Polymer Institute, University of Würzburg, 3Institute for Advanced Study, Technical University of Munich (TUM), 4George W Woodruff School of Mechanical Engineering, Georgia Institute of Technology This protocol serves as a comprehensive guideline to fabricate scaffolds via electrospinning with polymer melts in a direct writing mode. We systematically outline the process and define the appropriate parameter settings for achieving targeted scaffold architectures. Cancer Research Hyperpolarized 13C Metabolic Magnetic Resonance Spectroscopy and Imaging Eugen Kubala*1,2,3, Kim A. Muñoz-Álvarez*1, Geoffrey Topping1, Christian Hundshammer1,2, Benedikt Feuerecker1, Pedro A. Gómez3,4, Giorgio Pariani1,5,6, Franz Schilling1, Steffen J. Glaser2, Rolf F. Schulte3, Marion I. Menzel3, Markus Schwaiger1 1Department of Nuclear Medicine, Klinikum rechts der Isar, Technische Universität München, 2Department of Chemistry, Technische Universität München, 3GE Global Research, 4Zentralinstitut für Medizintechnik der Technischen Universität München (IMETUM), Technische Universität München, 5Institute for Biological and Medical Imaging (IBMI), Helmholtz Zentrum München, 6IDG Institute of Developmental Genetics, Helmholtz Zentrum München Dynamic nuclear polarization with subsequent sample dissolution has enabled real-time studies of metabolism in biological systems. Hyperpolarized [1-13C]pyruvate was used to study lactate dehydrogenase activity in a prostate carcinoma cell line in vitro. Medicine Tissue-simulating Phantoms for Assessing Potential Near-infrared Fluorescence Imaging Applications in Breast Cancer Surgery Rick Pleijhuis1, Arwin Timmermans1, Johannes De Jong1, Esther De Boer1, Vasilis Ntziachristos2, Gooitzen Van Dam1 1Department of Surgery, University Medical Center Groningen, 2Helmholtz Zentrum Munich, Technical University of Munich Near-infrared fluorescence (NIRF) imaging may improve therapeutic outcome of breast cancer surgery by enabling intraoperative tumor localization and evaluation of surgical margin status. Using tissue-simulating breast phantoms containing fluorescent tumor-simulating inclusions, potential clinical applications of NIRF imaging in breast cancer patients can be assessed for standardization and training purposes. Bioengineering Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology Xiaofeng Cui*1,2,3, Guifang Gao*2,4, Tomo Yonezawa5,6, Guohao Dai1 1Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 2Stemorgan Inc., 3Institute of Advanced Study, Technical University of Munich, 4Institute of Virology, School of Medicine, Wuhan University, 5Department of Molecular and Experimental Medicine, The Scripps Research Institute, 6Research Institute for Biomedical Sciences, Tokyo University of Science The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.