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University of Duisburg-Essen

6 ARTICLES PUBLISHED IN JoVE

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Biology

Chick ex ovo Culture and ex ovo CAM Assay: How it Really Works
Daniel S. Dohle 1, Susanne D. Pasa 1, Sebastian Gustmann 2, Markus Laub 3, Josef H. Wissler 4, Herbert P. Jennissen 1, Nicole Dünker 2
1Institute for Physiological Chemistry, Department of Biochemical Endocrinology, University of Duisburg-Essen, 2Institute for Anatomy, Department of Neuroanatomy, University of Duisburg-Essen, 3Morphoplant GmbH, 4ARCONS Institute for Applied Research and Didactics

The chick chorioallantoic membrane (CAM) is a unique, naturally immunodeficient supportive culture environment to study angiogenesis and tumorigenesis. This video article demonstrates the different steps in chick ex ovo culture, application of potentially angiogenic substances and successful inoculation of tumor cells and tissues on the surface of the CAM.

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Biology

In vivo-like Organotypic Murine Retinal Wholemount Culture
Sebastian Gustmann 1, Nicole Dünker 1
1Institute for Anatomy, Department of Neuroanatomy, University of Duisburg-Essen

This video article demonstrates the establishment of organotypic retinal wholemount cultures and a cytospin procedure for analysis of exogenously induced effects. Organotypic retinal wholemount cultures mimic the in vivo situation and significantly facilitate the accessibility of murine retinas for experimental manipulations while circumventing the disadvantages of classical murine animal models.

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Neuroscience

Intravascular Perfusion of Carbon Black Ink Allows Reliable Visualization of Cerebral Vessels
Mohammad R. Hasan 1, Josephine Herz 1, Dirk M. Hermann 1, Thorsten R. Doeppner 1
1Department of Neurology, University of Duisburg-Essen Medical School

Analysis of rodent cerebrovascular anatomy plays an important role in experimental stroke research. In this context, intravascular perfusion with colored latex has been considered as a standard tool for several years. However, this technique implies distinct technical limitations, which undermine its reproducibility. Here, we describe a simple method to visualize cerebral vessels in a reproducible manner. Injection of a mixture of two commercially available carbon black inks through the left myocardial ventricle results in adequate filling of cerebral vessels with high contrast visualization. We have successfully applied this technique to identify anastomotic points between cerebral vascular territories of mice with different genetic backgrounds. We finally give evidence that this novel and simple method for vessel staining can be combined with triphenyltetrazolium chloride (TTC) staining - a widely used tool to observe and analyze infarct volumes in mice.

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Biology

Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques
Nico Grüner 1, Oumaima Stambouli 1, R. Stefan Ross 1
1Institute of Virology, National Reference Centre for Hepatitis C, Essen University Hospital, University of Duisburg-Essen

The preparing and processing of dried blood spots (DBS) for their final analysis are still poorly standardized for most diagnostic applications. To overcome this shortcoming, a comprehensive step-by-step protocol is suggested and subsequently evaluated with regard to its effectiveness for detecting markers of viral infections.

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Neuroscience

Implantation of Miniosmotic Pumps and Delivery of Tract Tracers to Study Brain Reorganization in Pathophysiological Conditions
Eduardo H. Sanchez-Mendoza 1, Jeismar Carballo 1,2,3, Marines Longart 2, Dirk M. Hermann 1, Thorsten R. Doeppner 1
1Department of Neurology, University of Duisburg-Essen, 2Neuroscience Unit, Institute for Advanced Studies (IDEA), 3Department of Cell Biology, Simon Bolivar University

In order to study brain reorganization under pathological conditions we used miniosmotic pumps for direct protein delivery into the brain circumventing the blood brain barrier. Tract tracers are then injected to study alterations in brain connectivity under the influence of the protein.

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Cancer Research

Transfer of Manipulated Tumor-associated Neutrophils into Tumor-Bearing Mice to Study their Angiogenic Potential In Vivo
Ekaterina Pylaeva 1, Ilona Spyra 1, Sharareh Bordbari 1, Stephan Lang 1, Jadwiga Jablonska 1
1Department of Otorhinolaryngology, University Hospital, University of Duisburg-Essen

Here, we show therapeutic potential of anti-angiogenic tumor-associated neutrophils after their transfer into tumor-bearing mice. This protocol can be used to manipulate neutrophil activity ex vivo and to subsequently evaluate their functionality in vivo in developing tumors. It is an appropriate model for studying potential neutrophil-based immunotherapies.

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