Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin
2 ARTICLES PUBLISHED IN JoVE
Biology
Nuclei Isolation from Adult Mouse Kidney for Single-Nucleus RNA-Sequencing
Janna Leiz *1,2, Christian Hinze *1,2, Anastasiya Boltengagen 3, Caroline Braeuning 4, Christine Kocks 3, Nikolaus Rajewsky 3, Kai M. Schmidt-Ott 1,2
1Department of Nephrology and Medical Intensive Care Medicine, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 2Molecular and Translational Kidney Research, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 3Berlin Institute of Medical Systems Biology (BIMSB), Systems Biology of Gene Regulatory Elements, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 4Berlin Institute of Medical Systems Biology (BIMSB), Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC)
Here, we present a protocol to isolate high-quality nuclei from frozen mouse kidneys that improve the representation of medullary kidney cell types and avoids the gene expression artifacts from enzymatic tissue dissociation.
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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