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German Cancer Research Center

7 ARTICLES PUBLISHED IN JoVE

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Medicine

Quantitative Mass Spectrometric Profiling of Cancer-cell Proteomes Derived From Liquid and Solid Tumors
Hanibal Bohnenberger 1, Philipp Ströbel 1, Sebastian Mohr 2, Jasmin Corso 3, Tobias Berg 2, Henning Urlaub 3,4, Christof Lenz 3,4, Hubert Serve 2,5,6, Thomas Oellerich 2,5,6
1Institute of Pathology, University Medical Center, Göttingen, 2Department of Hematology/Oncology, Goethe University of Frankfurt, 3Bioanalytical Mass Spectrometry Group, Max Planck Institute for Biophysical Chemistry, 4Bioanalytics, Institute of Clinical Chemistry, University Medical Center, Göttingen, 5German Cancer Consortium, 6German Cancer Research Center

In-depth analyses of cancer cell proteomes facilitate identification of novel drug targets and diagnostic biomarkers. We describe an experimental workflow for quantitative analysis of (phospho-)proteomes in cancer cell subpopulations derived from liquid and solid tumors. This is achieved by combining cellular enrichment strategies with quantitative Super-SILAC-based mass spectrometry.

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Bioengineering

Atomic Force Microscopy Imaging and Force Spectroscopy of Supported Lipid Bilayers
Joseph D. Unsay 1,2,3, Katia Cosentino 1,2, Ana J. García-Sáez 1,2
1Interfaculty Institute for Biochemistry, 2Max Planck Institute for Intelligent Systems, 3German Cancer Research Center

We describe a protocol for preparation of supported lipid bilayers and its characterization using atomic force microscopy and force spectroscopy.

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Medicine

Isolation of Endothelial Progenitor Cells from Healthy Volunteers and Their Migratory Potential Influenced by Serum Samples After Cardiac Surgery
Christoph Emontzpohl 1,2, David Simons 3, Sandra Kraemer 4, Andreas Goetzenich 4, Gernot Marx 1, Jürgen Bernhagen 5,6, Christian Stoppe 1
1Department of Intensive Care Medicine, University Hospital Aachen, 2Institute of Biochemistry and Molecular Biology, University Hospital Aachen, 3Department of Radiology, German Cancer Research Center, 4Department of Thoracic and Cardiovascular Surgery, University Hospital Aachen, 5Department of Vascular Biology, Institute for Stroke and Dementia Research (ISD), Klinikum der Universität München, 6Deutsches Zentrum für Herz-/Kreislaufkrankheiten (DZHK), Munich Heart Alliance

Endothelial progenitor cells (EPCs) are crucially involved in the neovascularization of ischemic tissues. This method describes the isolation of human EPCs from peripheral blood, as well as the identification of their migratory potential against serum samples of cardiac surgical patients.

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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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Cancer Research

Machine Learning Algorithms for Early Detection of Bone Metastases in an Experimental Rat Model
Stephan Ellmann 1, Lisa Seyler 1, Clarissa Gillmann 2, Vanessa Popp 1, Christoph Treutlein 1, Aline Bozec 3, Michael Uder 1, Tobias Bäuerle 1
1Department of Radiology, University Hospital Erlangen, Friedrich-Alexander Universität Erlangen-Nürnberg, 2Department of Medical Physics in Radiation Oncology, German Cancer Research Center, 3Department of Internal Medicine 3, University Hospital Erlangen, Friedrich-Alexander Universität Erlangen-Nürnberg

This protocol was designed to train a machine learning algorithm to use a combination of imaging parameters derived from magnetic resonance imaging (MRI) and positron emission tomography/computed tomography (PET/CT) in a rat model of breast cancer bone metastases to detect early metastatic disease and predict subsequent progression to macrometastases.

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Cancer Research

Mast Cells in the Microenvironment of Hepatocellular Carcinoma Confer Favorable Prognosis: A Retrospective Study using QuPath Image Analysis Software
Esraa Ali 1, Lenka Červenková 2,3, Richard Pálek 2,4, Filip Ambrozkiewicz 1, Sergii Pavlov 1, Wenjing Ye 1, Petr Hošek 2, Ondrej Daum 5,6, Václav Liška 2,4, Kari Hemminki 1,7, Andriy Trailin 1
1Laboratory of Translational Cancer Genomics, Biomedical Center, Faculty of Medicine in Pilsen, Charles University, 2Laboratory of Cancer Treatment and Tissue Regeneration, Biomedical Center, Faculty of Medicine in Pilsen, Charles University, 3Department of Pathology, Third Faculty of Medicine, Charles University, 4Department of Surgery and Biomedical Center, Faculty of Medicine in Pilsen, Charles University, 5Sikl's Institute of Pathology, Faculty of Medicine and Teaching Hospital in Pilsen, Charles University, 6Bioptická Laboratoř s.r.o., 7Department of Cancer Epidemiology, German Cancer Research Center

The presence of mast cells in the inner margin and peritumor areas of hepatocellular carcinoma after resection confers a favorable prognosis. This study endorses QuPath image analysis software as a promising platform that could meet the need for reproducibility, consistency, and accuracy in digital pathology.

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Immunology and Infection

Operating and Biocontainment Procedures of a Facility for Laboratory Mice with a Natural Microbiome: Immunophenotyping Procedure
Natascha Drude 1, Kai Diederich 2, Claudia U. Duerr 3, Nadine Haase 4, Christoph Harms 5,6,7,8, Frank Heppner 9,10,11, Marina Jendrach 9,10,11, Pia Kahnau 2, Marina Kolesnichenko 12, Lars Lewejohann 2,13, Claudia Kurreck 1, Anke Lohan 14, Marcus A. Mall 15,16, Dominik Müller 4, Stefan Nagel-Riedasch 14, Bastian Opitz 17, Laura Schaupp 15,16, Gilbert Schönfelder 2,18, Amelie Weber 3, Gerald Willimsky 19,20,21, Yalei Zang 1,22, Stephan P. Rosshart *23,24, Andreas Diefenbach *3, Stefan Jordan *3
1BIH QUEST Center for Responsible Research, Berlin Institute of Health (BIH) at Charité - Universitätsmedizin Berlin, 2German Federal Institute for Risk Assessment (BfR), German Centre for the Protection of Laboratory Animals (Bf3R), 3Institute of Microbiology, Infectious Diseases and Immunology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 4Experimental and Clinical Research Center (ECRC) & Max-Delbrück-Centrum für Molekulare Medizin in der Helmholtz-Gemeinschaft (MDC), 5Department of Experimental Neurology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Klinik und Hochschulambulanz für Neurologie, 6Center for Stroke Research Berlin, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 7Einstein Center for Neuroscience, 8German Center for Cardiovascular Research (DZHK), partner site Berlin, 9Department of Neuropathology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 10Cluster of Excellence, NeuroCure, 11German Center for Neurodegenerative Diseases (DZNE) Berlin, 12Department of Hepatology and Gastroenterology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 13Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Freie Universität Berlin, 14Forschungseinrichtungen für Experimentelle Medizin/FEM, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 15Department of Pediatric Respiratory Medicine, Immunology and Critical Care Medicine, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 16German Center for Lung Research (DZL), associated partner site, Berlin, 17Department of Infectious Diseases, Respiratory Medicine and Critical Care, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 18Division of Toxicology, Institute of Clinical Pharmacology and Toxicology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 19Institute of Immunology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 20German Cancer Research Center, 21German Cancer Consortium, partner site Berlin, 22Department of Radiology (including Pediatric Radiology), Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 23Department of Microbiome Research, University Hospital Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), 24Department of Medicine II, Faculty of Medicine, Medical Center - University of Freiburg

Here, we describe the structure and operating procedures, including microbial containment measures of a facility for "Wilding mice" using blood sampling for immunophenotyping as an example.

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