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Medical University Innsbruck

15 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Orthotopic Hind-Limb Transplantation in Rats
Robert Sucher *1, Rupert Oberhuber *1, Christian Margreiter 1, Guido Rumberg 1, Rishi Jindal 2, WP Andrew Lee 2, Raimund Margreiter 1, Johann Pratschke 1, Stefan Schneeberger 1, Gerald Brandacher 1
1Department of Visceral, Transplant, and Thoracic Surgery, Daniel Swarovski Research Laboratory, Innsbruck Medical University, 2Department of Surgery, Division of Plastic and Reconstructive Surgery, University of Pittsburgh Medical Center

Here we describe the orthotopic rat hind-limb transplantation procedure, which seems to be the gold standard in vivo model for composite tissue allotransplantation research.

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Medicine

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation (VCA) Research
Zuhaib Ibrahim 1, Damon S. Cooney 1, Jaimie T. Shores 1, Justin M. Sacks 1, Eric G. Wimmers 1, Steven C. Bonawitz 1, Chad Gordon 1, Dawn Ruben 1, Stefan Schneeberger 1, W. P. Andrew Lee 1, Gerald Brandacher 1
1Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine

Vascularized Composite Allotransplantations (VCA) have become a clinical reality. However, broad clinical application of VCA is limited by chronic multi-drug immunosuppression. The authors present a reliable and reproducible large animal model to translate novel immunomodulatory strategies that can minimize or potentially eliminate the need of immunosuppression in VCA.

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Medicine

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique
Rupert Oberhuber *1, Benno Cardini *1, Markus Kofler 1, Paul Ritschl 1, Robert Oellinger 1, Felix Aigner 1, Robert Sucher 1, Stefan Schneeberger 1, Johann Pratschke 1, Gerald Brandacher 2, Manuel Maglione 1
1Center of Operative Medicine, Department of Visceral, Transplant and Thoracic Surgery, Innsbruck Medical University, 2Department of Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.

Herein, we provide additional details of this technique to supplement the video.

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JoVE Journal

Shock Wave Application to Cell Cultures
Johannes Holfeld 1, Can Tepeköylü 1, Radoslaw Kozaryn 1, Wolfgang Mathes 1, Michael Grimm 1, Patrick Paulus 2
1Department of Cardiac Surgery, Innsbruck Medical University, 2Clinic of Anesthesiology, Intensive Care Medicine and Pain Therapy, Goethe-University Hospital

Shock waves nowadays are well known for their regenerative effects. Therefore in vitro experiments are of increasing interest. We therefore developed a model for in vitro shock wave trials (IVSWT) that enables us to mimic in vivo conditions thereby avoiding distracting physical effects.

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Medicine

A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice
Byoungchol Oh *1, Georg J. Furtmüller *1, Michael Sosin *1, Madeline L. Fryer 1, Lawrence J. Gottlieb 2,3, Michael R. Christy 4, Gerald Brandacher 1,5,6, Amir H. Dorafshar 1,5,6
1Johns Hopkins University School of Medicine, 2Burn and Complex Wound Center, 3Section of Plastic and Reconstructive Surgery, University of Chicago Medical Center, 4Division of Plastic, Reconstructive, and Maxillofacial Surgery, R Adams Cowley Shock Trauma Center, 5Department of Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine, 6Vascularized Composite Allotransplantation (VCA) Lab, Johns Hopkins University School of Medicine

To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

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Medicine

Orthotopic Hind Limb Transplantation in the Mouse
Georg J. Furtmüller *1, Byoungchol Oh *1, Johanna Grahammer *2, Cheng-Hung Lin 3, Robert Sucher 4, Madeline L. Fryer 1, Giorgio Raimondi 1, W.P. Andrew Lee 1, Gerald Brandacher 1
1Department of Plastic and Reconstructive Surgery, Vascularized Composite Allotransplantation (VCA) Laboratory, Johns Hopkins University School of Medicine, 2Department of Visceral, Transplant and Thoracic Surgery, Innsbruck Medical University, 3Center for Vascularized Composite Allotransplantation, Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital and School of Medicine, 4Department of General, Visceral and Transplant Surgery, Charite Berlin

This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA).

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Medicine

Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique
Benno Cardini *1, Rupert Oberhuber *1, Sven R Hein 1, Rebecca Eiter 1, Martin Hermann 2, Markus Kofler 1,3, Stefan Schneeberger 1, Gerald Brandacher 1,4, Manuel Maglione 1
1Center of Operative Medicine, Department of Visceral, Transplant and Thoracic Surgery, Medical University Innsbruck, 2Department of Anesthesiology and Critical Care Medicine, Medical University Innsbruck, 3Department of Cardiac Surgery, Medical University Innsbruck, 4Department of Plastic and Reconstructive Surgery, Vascularized Composite Allotransplantation (VCA) Laboratory, Johns Hopkins University School of Medicine

Among abdominal solid organ transplantation, pancreatic grafts are prone to develop severe ischemia reperfusion injury-associated graft damage, leading eventually to early graft loss. This protocol describes a model of murine pancreas transplantation using a non-suture cuff technique, ideally suited for analyzing these early, deleterious damages.

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Immunology and Infection

Murine Full-thickness Skin Transplantation
Chih-Hsien Cheng *1,2, Chen-Fang Lee *1,2, Madeline Fryer *3, Georg J. Furtmüller 3, Byoungchol Oh 3, Jonathan D. Powell 1, Gerald Brandacher 3
1Sidney-Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, 2Department of Liver and Transplantation Surgery, Chang-Gung Transplantation Institute, Chang-Gung Memorial Hospital, Chang-Gung University College of Medicine, 3Vascularized Composite Allotransplantation Laboratory, Department of Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine

Murine full-thickness skin transplantation is a well-established model to study rejection in an alloimmune setting. Here, we provide a tutorial of each step involved in performing a BALB/c-->C57BL/6 full-thickness skin transplant.

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Cancer Research

Harnessing the DNA Dye-triggered Side Population Phenotype to Detect and Purify Cancer Stem Cells from Biological Samples
Maximilian Boesch *1,2,3, Elisabeth Hoflehner 2,3, Dominik Wolf 2,3,4, Guenther Gastl 2, Sieghart Sopper *2,3
1Institute of Immunobiology, Kantonsspital St. Gallen, 2Internal Medicine V, Medical University of Innsbruck, 3Tyrolean Cancer Research Institute (TKFI), 4Medical Clinic III, Oncology, Hematology, Immunoncology and Rheumatology, University Clinic Bonn (UKB)

Methods allowing the characterization and isolation of stem cell populations from biological samples are critical for the advance of stem cell-targeted treatments in cancer and beyond. Here, we provide a detailed protocol for cancer stem cell isolation using the dye-triggered side population phenotype.

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Developmental Biology

Detection of Residual Donor Erythroid Progenitor Cells after Hematopoietic Stem Cell Transplantation for Patients with Hemoglobinopathies
Roman Crazzolara 1, Gabriele Kropshofer 1, Michael Steurer 2, Sieghart Sopper 2,3, Wolfgang Schwinger 4
1Department of Pediatrics, Medical University Innsbruck, 2Department of Internal Medicine V (Hematology & Oncology), Medical University Innsbruck, 3Tyrolean Cancer Research Institute, 4Department of Pediatrics, Medical University Graz

Quantification of donor-derived cells is required to monitor engraftment after stem cell transplantation in patients with hemoglobinopathies. A combination of flow cytometry-based cell sorting, colony formation assay, and subsequent analysis of short tandem repeats may be used to assess the proliferation and differentiation of progenitors in the erythroid compartment.

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Cancer Research

A Data Integration Workflow to Identify Drug Combinations Targeting Synthetic Lethal Interactions
Maximilian Marhold 1, Andreas Heinzel 2, Almas Merchant 1, Paul Perco 3, Michael Krainer 1
1Department of Internal Medicine I - Division of Oncology, Comprehensive Cancer Center, Medical University of Vienna, 2Department of Nephrology, Medical University of Vienna, 3Department of Internal Medicine IV, Medical University Innsbruck

Large genetic screens in model organisms have led to the identification of negative genetic interactions. Here, we describe a data integration workflow using data from genetic screens in model organisms to delineate drug combinations targeting synthetic lethal interactions in cancer.

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Neuroscience

Brain Death Induction in Mice Using Intra-Arterial Blood Pressure Monitoring and Ventilation via Tracheostomy
Paul V. Ritschl *1,2,3, Lena Hofhansel *2,4, Bernhard Flörchinger 5, Rupert Oberhuber 2, Robert Öllinger 1, Johann Pratschke 1, Katja Kotsch 6
1Department of Surgery Campus Charité Mitte/Campus Virchow-Klinikum, Charité - Universitätsmedizin Berlin, 2Department of Visceral, Transplant and Thoracic Surgery, Medical University of Innsbruck, 3Charité Clinician Scientist Program, Berlin Institute of Health, 4Department of Psychiatry, Psychotherapy and Psychosomatics, RWTH Aachen University, 5Department of Cardiothoracic Surgery, University Medical Center Regensburg, 6Department of General, Visceral and Vascular Surgery, Charité - Universitätsmedizin Berlin

We present a murine model of brain death induction in order to evaluate the influence of its pathophysiological effects on organs as well as on consecutive grafts in the context of solid organ transplantation.

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Biology

Exosome Isolation after in vitro Shock Wave Therapy
Leo Pölzl 1,2, Felix Nägele 1, Jakob Hirsch 1, Michael Graber 1, Michael Grimm 1, Can Gollmann-Tepeköylü 1, Johannes Holfeld 1
1Department of Cardiac Surgery, Medical University Innsbruck, 2Institute of Clinical and Functional Anatomy, Innsbruck Medical University

Exosomes are released upon in vitro application of shockwaves. Here, we describe how to apply shockwaves on cultured endothelial cells and subsequently isolate exosomes for further investigation.

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Medicine

Performing Repeated Intraoperative Impedance Telemetry Measurements during Cochlear Implantation
Nora M. Weiss 1,2,3,4,5, Stefan Hans 6, Martin Wozniak 6, Aline Föger 6, Stefan Dazert 1, Vincent Van Rompaey 2,7, Paul Van de Heyning 2,7, Joachim Schmutzhard 8, Angelika Dierker 6
1Department of Otorhinolaryngology-Head and Neck Surgery, Ruhr-University Bochum, St. Elisabeth-Hospital Bochum, 2Department of Translational Neurosciences, Faculty of Medicine and Health Sciences, University of Antwerp, 3International Graduate School of Neuroscience (IGSN), Ruhr-University Bochum, 4Technical University of Munich, 5Department of Otorhinolaryngology, TUM School of Medicine, Klinikum rechts der Isar, 6MED-EL, 7Department of Otorhinolaryngology and Head & Neck Surgery, Antwerp University Hospital, 8Department of Otorhinolaryngology-Head and Neck Surgery, Medical University Innsbruck

Here we present a protocol to conduct repeated impedance telemetry measurements during cochlear implantation (CI). They may allow conclusions on the electrode's and implant's function. Repeated impedance measurements enable objective feedback on whether the electrode is positioned inside the perilymph or outside the inner ear.

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Medicine

Electrically Evoked Stapedius Reflex Measurements in Cochlear Implantation and Its Application in the Postoperative Fitting Process
Maria Magdalena Mair 1, Ruth Schröcksnadel 1, Kurt Stephan 2, Josef Seebacher 2, Philipp Zelger 2, Franz Muigg 1, Timo Gottfried 3, Annett Franke-Trieger 4, Joachim Schmutzhard 3
1Department for Hearing, Speech and Voice Disorders, Tirol-Kliniken GmbH, 2Department for Hearing, Speech and Voice Disorders, Medical University of Innsbruck, 3Department of Otorhinolaryngology, Medical University Innsbruck, 4Saxonian Cochlear Implant Centre, Department of Otorhinolaryngology, Head and Neck Surgery, Faculty of Medicine (and University Hospital) Carl Gustav Carus, Technical University of Dresden

The present protocol describes the measurement of the electrically evoked stapedius reflex (eSR) via cochlear implant (CI). Two applications are discussed: intraoperative detection of eSR for verification of the coupling between the cochlear implant and the auditory nerve and postoperative measurement of eSR thresholds (eSRT) for CI fitting.

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