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Heidelberg University Hospital

11 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

An Experimental Model to Study Tuberculosis-Malaria Coinfection upon Natural Transmission of Mycobacterium tuberculosis and Plasmodium berghei
Ann-Kristin Mueller 1, Jochen Behrends 2, Jannike Blank 2, Ulrich E. Schaible 2, Bianca E. Schneider 2
1Department of Infectious Diseases, Parasitology Unit, University Hospital Heidelberg, 2Priority Area Infections, Research Center Borstel

Tuberculosis and malaria are two of the most prevalent infections in humans and major causes of morbidity and mortality in impoverished populations in the tropics. We established an experimental model system to study outcome of malaria-tuberculosis coinfection in mice after challenge with both pathogens via their natural route of infection.

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Medicine

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts
Shaoping Hou 1, Armin Blesch 2, Paul Lu 3,4
1Spinal Cord Research Center, Department of Neurobiology & Anatomy, Drexel University College of Medicine, 2Spinal Cord Injury Center, Heidelberg University Hospital, 3Veterans Administration Medical Center, San Diego, CA, 4Department of Neurosciences, University of California, San Diego

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

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Medicine

In Vivo Tracking of Edema Development and Microvascular Pathology in a Model of Experimental Cerebral Malaria Using Magnetic Resonance Imaging
Angelika Hoffmann 1,2, Xavier Helluy 2,3, Manuel Fischer 2, Ann-Kristin Mueller 4,5, Sabine Heiland 2, Mirko Pham 1,6, Martin Bendszus 1, Johannes Pfeil 4,5,7
1Department of Neuroradiology, Heidelberg University Hospital, 2Division of Experimental Radiology, Department of Neuroradiology, Heidelberg University Hospital, 3NeuroImaging Centre Research, Department of Neuroscience, Ruhr-University Bochum, 4Centre for Infectious Diseases, Parasitology Unit, Heidelberg University Hospital, 5German Centre for Infection Research (DZIF), 6Department of Neuroradiology, University of Würzburg, 7Center for Childhood and Adolescent Medicine, General Pediatrics, Heidelberg University Hospital

We describe a mouse model of experimental cerebral malaria and show how inflammatory and microvascular pathology can be tracked in vivo using magnetic resonance imaging.

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Developmental Biology

CRISPR-mediated Loss of Function Analysis in Cerebellar Granule Cells Using In Utero Electroporation-based Gene Transfer
Weijun Feng *1, Lena Herbst *2, Peter Lichter 2, Stefan M. Pfister 3,4,5, Hai-Kun Liu 1, Daisuke Kawauchi 3,4
1Division of Molecular Neurogenetics, German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, 2Division of Molecular Genetics, German Cancer Research Center (DKFZ), and German Cancer Consortium (DKTK), 3Hopp-Children's Cancer Center at the NCT Heidelberg (KiTZ), 4Division of Pediatric Neurooncology, German Cancer Research Center (DKFZ), and German Cancer Consortium (DKTK), 5Department of Pediatric Hematology and Oncology, Heidelberg University Hospital

Conventional loss-of-function studies of genes using knockout animals have often been costly and time-consuming. Electroporation-based CRISPR-mediated somatic mutagenesis is a powerful tool to understand gene functions in vivo. Here, we report a method to analyze knockout phenotypes in proliferating cells of the cerebellum.

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Engineering

Protocol for the Evaluation of MRI Artifacts Caused by Metal Implants to Assess the Suitability of Implants and the Vulnerability of Pulse Sequences
Tim Hilgenfeld 1, Marcel Prager 1, Franz S. Schwindling 2, Johann M.E. Jende 1, Peter Rammelsberg 2, Martin Bendszus 1, Sabine Heiland 1, Alexander Juerchott 1
1Department of Neuroradiology, Heidelberg University Hospital, 2Department of Prosthodontics, Heidelberg University Hospital

We describe a standardized method to evaluate magnetic resonance imaging artifacts caused by implants to estimate the suitability of the implants for magnetic resonance imaging and/or the vulnerability of different pulse sequences to metallic artifacts simultaneously.

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Immunology and Infection

Detection of Low Copy Number Integrated Viral DNA Formed by In Vitro Hepatitis B Infection
Thomas Tu 1,2, Stephan Urban 1,3
1Department of Infectious Diseases, Molecular Virology, Heidelberg University Hospital, 2Gastroenterology and Liver Research Laboratory, Ingham Institute, 3German Center for Infection Research (DZIF)

We describe here the in vitro generation of HBV DNA via a Hepatitis B virus infection system and the highly sensitive detection of its (1–2 copies) integration using inverse nested PCR.

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Cancer Research

Cell Cycle-specific Measurement of γH2AX and Apoptosis After Genotoxic Stress by Flow Cytometry
Ramon Lopez Perez 1,2, Franziska Münz 1,2, Jonas Kroschke 1,2, Jannek Brauer 1,2, Nils H. Nicolay 1,2,3, Peter E. Huber 1,2
1CCU Molecular and Radiation Oncology, German Cancer Research Center, 2Department of Radiation Oncology, Heidelberg University Hospital, 3Department of Radiation Oncology, Freiburg University Medical Center

The presented method combines the quantitative analysis of DNA double-strand breaks (DSBs), cell cycle distribution and apoptosis to enable cell cycle-specific evaluation of DSB induction and repair as well as the consequences of repair failure.

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JoVE Core

Flow Cytometry Analysis of Immune Cell Subsets within the Murine Spleen, Bone Marrow, Lymph Nodes and Synovial Tissue in an Osteoarthritis Model
Patrick Haubruck 1,2, Aimee C. Colbath 1,3, Yolanda Liu 1, Shihani Stoner 1, Cindy Shu 1, Christopher B. Little 1
1Raymond Purves Bone and Joint Research Laboratory, Institute of Bone and Joint Research, Kolling Institute, Royal North Shore Hospital, University of Sydney, 2HTRG - Heidelberg Trauma Research Group, Center for Orthopedics, Trauma Surgery and Spinal Cord Injury, Trauma and Reconstructive Surgery, Heidelberg University Hospital, 3Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University

Here, we describe a detailed and reproducible flow cytometry protocol to identify monocyte/macrophage and T-cell subsets using both extra- and intracellular staining assays within the murine spleen, bone marrow, lymph nodes and synovial tissue, utilizing an established surgical model of murine osteoarthritis.

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Immunology and Infection

Adapting Gastrointestinal Organoids for Pathogen Infection and Single Cell Sequencing under Biosafety Level 3 (BSL-3) Conditions
Megan L. Stanifer 1,2, Steeve Boulant 2,3,4
1Department of Infectious Diseases, Molecular Virology, Heidelberg University Hospital, 2Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, 3Department of Infectious Diseases, Virology, Heidelberg University Hospital, 4Research Group “Cellular Polarity and Viral Infection”, German Cancer Research Center (DKFZ)

This protocol describes how to infect human intestinal organoids from either their apical or basolateral side to characterize host/pathogen interactions at the single-cell level using single-cell RNA sequencing (scRNAseq) technology.

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Medicine

Endotracheal Intubation via Tracheotomy and Subsequent Thoracotomy in Rats for Non-Survival Applications
Alexander Studier-Fischer 1,2,3,4, Berkin Özdemir 1,3,4, Karl-Friedrich Kowalewski 2,3,4, Caelan Max Haney 2,3,4,5, Maurice Stephan Michel 2,3,4, Gabriel Alexander Salg 1, Franck Billmann 1
1Department of General, Visceral, and Transplantation Surgery, Heidelberg University Hospital, 2Department of Urology and Urosurgery, University Medical Center Mannheim, Medical Faculty of the University of Heidelberg, 3Intelligent Systems and Robotics in Urology (ISRU), German Cancer Research Center (DKFZ) Heidelberg, 4DKFZ Hector Cancer Institute, University Medical Center Mannheim, 5Department of Urology, University Hospital Leipzig

Here we introduce a standardized procedure for endotracheal intubation via tracheotomy followed by thoracotomy in rats, aimed at enhancing the precision and reproducibility of non-survival applications requiring invasive ventilation and exposure of thoracic organs in in vivo rat models.

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Medicine

Controlled Reversible Visceral Arterial Ischemia, Venous Congestion and Combined Malperfusion via Midline Laparotomy in Rats
Alexander Studier-Fischer 1,2,3,4, Berkin Özdemir 1,3,4, Karl-Friedrich Kowalewski 2,3,4, Caelan Max Haney 2,3,4,5, Luisa Egen 2,3,4, Maurice Stephan Michel 2,3,4, Markus Mieth 1, Zoltan Czigany 1, Gabriel Alexander Salg 1, Henrik Nienhüser 1, Franck Billmann 1
1Department of General, Visceral and Transplantation Surgery, Heidelberg University Hospital, 2Department of Urology and Urosurgery, University Medical Center Mannheim, Medical Faculty of the University of Heidelberg, 3Division of Intelligent Systems and Robotics in Urology (ISRU), German Cancer Research Center (DKFZ) Heidelberg, 4DKFZ Hector Cancer Institute at the University Medical Center Mannheim, 5Department of Urology, University Hospital Leipzig

This article introduces a standardized procedure for controlled, reversible malperfusion of visceral organs in rat models. The aim is to induce these malperfusion states with a high degree of methodological certainty and control while maintaining technical simplicity and error resilience.

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