University of Munich

4 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Assessment of Endothelial Cell Migration After Exposure to Toxic Chemicals

Dirk Steinritz^1,2, Annette Schmidt^1,3, Frank Balszuweit¹, Horst Thiermann¹, Marwa Ibrahim³, Birgit Bölck³, Wilhelm Bloch³

¹Bundeswehr Institute of Pharmacology and Toxicology, ²Walther Straub Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universität München, ³Department of Molecular and Cellular Sports Medicine, German Sports University Cologne

Investigation of early endothelial cell (EEC) migration is important to understand the pathophysiology of certain illnesses and to potentially identify novel strategies for therapeutic intervention. The following protocol describes techniques to assess cell migration that have been adapted for the investigation of EEC.

Genetics

A Customizable Protocol for String Assembly gRNA Cloning (STAgR)

Christopher T. Breunig^1,2, Andrea M. Neuner^1,2, Jessica Giehrl-Schwab³, Wolfgang Wurst³, Magdalena Götz^2,4, Stefan H. Stricker^1,2

¹MCN Junior Research Group, Munich Center for Neurosciences, Ludwig Maximilian Universitat, BioMedical Center, ²Institute of Stem Cell Research, Helmholtz Zentrum, German Research Center for Environmental Health, ³Institute of Developmental Genetics, Helmholtz Zentrum, German Research Center for Environmental Health, ⁴Physiological Genomics, Ludwig Maximilian Universitat, BioMedical Center

Here, we present string assembly gRNA cloning (STAgR), a method to easily multiplex gRNA vectors for CRISPR/Cas9 approaches. STAgR makes gRNA multiplexing simple, efficient and customizable.

Medicine

Assessment of the Acute Inhalation Toxicity of Airborne Particles by Exposing Cultivated Human Lung Cells at the Air-Liquid Interface

Amelie Tsoutsoulopoulos¹, Katrin Gohlsch², Niklas Möhle³, Andreas Breit², Sebastian Hoffmann⁵, Olaf Krischenowski^3,4, Harald Mückter², Thomas Gudermann², Horst Thiermann¹, Michaela Aufderheide^3,4, Dirk Steinritz^1,2

¹Bundeswehr Institute of Pharmacology and Toxicology, ²Walther Straub Institute of Pharmacology and Toxicology, University of Munich, ³Cultex Laboratories GmbH, ⁴Cultex Technologies GmbH (formerly Cultex Laboratories GmbH), ⁵seh consulting + services

We present a robust, transferable and predictive in vitro exposure system for the screening and monitoring of airborne particles concerning their acute pulmonary cytotoxicity by exposing cultivated human lung cells at the air-liquid interface (ALI).

Neuroscience

Cryo-section Dissection of the Adult Subependymal Zone for Accurate and Deep Quantitative Proteome Analysis

Christian Friess¹, Magdalena Götz^1,2,3, Jacob Kjell^1,2,4

¹Division of Physiological Genomics, Biomedical Center, Ludwig Maximilian University of Munich, ²Institute for Stem Cell Research, Helmholtz Zentrum München, ³SYNERGY, Excellence Cluster Systems Neurology, University of Munich, ⁴Department of Clinical Neuroscience, Karolinska Institutet

Cryo-section-dissection allows fresh, frozen preparation of the largest neurogenic niche in the murine brain for deep quantitative proteome analysis. The method is precise, efficient, and causes minimal tissue perturbation. Therefore, it is ideally suited for studying the molecular microenvironment of this niche, as well as other organs, regions, and species.