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University Hospital Tübingen

16 ARTICLES PUBLISHED IN JoVE

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Clinical Skills

COVID-19 / Coronavirus Outbreak: Protecting The Airway - Endotracheal Intubation
Robert Wunderlich 1, Eric Stricker 2, Almut Goll 1, Periklis Spiros Nikomanis 3, Helene Häberle 1, Alexander Münch 1
1Department of Anesthesiology and Intensive Care Medicine, University Clinic Tübingen, 2Center for Patient Safety and Simulation Tüpass, University Hospital Tübingen, 3Department of Anesthesiology and Intensive Care Medicine, Gesundbrunnen Hospital, SLK Clinical Centers

COVID-19 / Coronavirus Outbreak: Protecting The Airway - Endotracheal Intubation

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Biology

In Vitro Synthesis of Modified mRNA for Induction of Protein Expression in Human Cells
Meltem Avci-Adali 1, Andreas Behring 1, Heidrun Steinle 1, Timea Keller 1, Stefanie Krajeweski 1, Christian Schlensak 1, Hans P. Wendel 1
1Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen

In this video article, we describe the in vitro synthesis of modified mRNA for induction of protein expression in cells.

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Medicine

Evaluation of Stem Cell Properties in Human Ovarian Carcinoma Cells Using Multi and Single Cell-based Spheres Assays
Hui Wang 1,2, Anna Paczulla 1, Claudia Lengerke 1,2
1Department of Biomedicine, University Hospital Basel, 2Department of Internal Medicine II, University Hospital Tübingen

In vitro spheres assays are commonly used to identify cancer stem cells. Here we compare single with multi cell-based spheres assays. The more laborious single cell-based assays or methylcellulose supplementation give more accurate results while multi cell-based assays performed in liquid medium can be highly influenced by cell density.

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Behavior

How to Study Placebo Responses in Motion Sickness with a Rotation Chair Paradigm in Healthy Participants
Katja Weimer 1, Björn Horing 2, Eric R. Muth 2, Paul Enck 1
1Psychosomatic Medicine and Psychotherapy, University Hospital Tübingen, 2Department of Psychology, Clemson University

An experimental rotation chair paradigm is used to investigate whether a placebo intervention on motion sickness affects subjective outcome measures only or also affects behavioral and objective measures in a balanced placebo design.

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Immunology and Infection

A RAPID Method for Blood Processing to Increase the Yield of Plasma Peptide Levels in Human Blood
Pauline Teuffel 1, Miriam Goebel-Stengel 1,2, Tobias Hofmann 1, Philip Prinz 1, Sophie Scharner 1, Jan L. Körner 3, Carsten Grötzinger 3, Matthias Rose 1, Burghard F. Klapp 1, Andreas Stengel 1
1Charité Center for Internal Medicine and Dermatology, Division General Internal and Psychosomatic Medicine, Charité Universitätsmedizin Berlin, Campus Benjamin Franklin, 2Department of Internal Medicine, Institute of Neurogastroenterology and Motility, Martin-Luther Hospital, Academic Teaching Institution of Charité Universitätsmedizin Berlin, 3Department of Hepatology and Gastroenterology, Molecular Cancer Research Center (MKFZ), Campus Virchow-Klinikum, Charité-Universitätsmedizin Berlin

The RAPID blood processing method can be used in humans and yields higher peptide levels as well as allows for assessment of the correct molecular form. Therefore, this method will be a valuable tool in peptide research.

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Medicine

Induction of Nephrotic Syndrome in Mice by Retrobulbar Injection of Doxorubicin and Prevention of Volume Retention by Sustained Release Aprotinin
Bernhard N. Bohnert 1,2,3, Ferruh Artunc 1,2,3
1Department of Internal Medicine, Division of Endocrinology, Diabetology, Vascular Disease, Nephrology and Clinical Chemistry, University Hospital Tübingen, 2Institute of Diabetes Research and Metabolic Diseases (IDM) of the Helmholtz Center Munich, University Tübingen, 3German Center for Diabetes Research (DZD), University Tübingen

Here, we describe the induction of experimental nephrotic syndrome in 129S1/SvImJ mice by rapid retrobulbar injection of doxorubicin. We also treat nephrotic mice with sustained release pellets containing aprotinin to inhibit urinary serine protease activity and prevent sodium retention.

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Medicine

A Porcine Corneal Endothelial Organ Culture Model Using Split Corneal Buttons
Daniel A. Wenzel 1, Berenike C. Kunzmann 2, Nils A. Steinhorst 1, Martin S. Spitzer 1, Maximilian Schultheiss 1
1Department of Ophthalmology, University Medical Center Hamburg-Eppendorf (UKE), 2University Eye Hospital, Center For Ophthalmology, University Hospital Tübingen

Here, a step-by-step protocol for the preparation and cultivation of porcine split corneal buttons is presented. As this organo-typically cultivated organ culture model shows cell death rates within 15 days, comparable to human donor corneas, it represents the first model allowing long-term cultivation of non-human corneas without adding toxic dextran.

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Medicine

Sucrose Preference and Novelty-Induced Hypophagia Tests in Rats using an Automated Food Intake Monitoring System
Martha Anna Schalla *1, Stephanie Gladys Kühne *1, Tiemo Friedrich 1, Vivien Hanel 1, Peter Kobelt 1, Miriam Goebel-Stengel 1,2,3, Matthias Rose 1,4, Andreas Stengel 1,3
1Charité Center for Internal Medicine and Dermatology, Department for Psychosomatic Medicine, Charité-Universitätsmedizin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin and Berlin Institute of Health, 2Department of Internal Medicine, Helios Clinic, 3Department of Psychosomatic Medicine and Psychotherapy, University Hospital Tübingen, 4Department of Quantitative Health Sciences, University of Massachusetts Medical School

Presented here is a protocol to study depression-like and anhedonic behavior in rats. It combines two well-established behavioral methods, the sucrose preference and novelty-induced hypophagia tests, with an automated food and liquid intake monitoring system, to indirectly investigate rodent behavior using surrogate parameters.

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Bioengineering

Application of Atomic Force Microscopy to Detect Early Osteoarthritis
Marina Danalache *1, Aadhya Tiwari *1, Viktor Sigwart 1,2, Ulf Krister Hofmann 1,3
1Laboratory of Cell Biology, Department of Orthopaedic Surgery, University Hospital of Tübingen, 2Medical faculty of the University of Tübingen, 3Department of Orthopaedic Surgery, University Hospital of Tübingen

We present a method to investigate early osteoarthritic changes at the cellular level in articular cartilage by using atomic force microscopy (AFM).

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Neuroscience

Visualizing the Calcitonin Gene-Related Peptide Immunoreactive Innervation of the Rat Cranial Dura Mater with Immunofluorescence and Neural Tracing
Jia Wang *1, Dongsheng Xu *1, Jingjing Cui 1, Chen She 1, Hui Wang 1, Shuang Wu 1, Ling Zou 1, Jianliang Zhang 1, Wanzhu Bai 1
1Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences

Here we present a protocol to visualize spatial correlation of calcitonin gene-related peptide (CGRP)-immunoreactive nerve fibers and blood vessels in the cranial dura mater using immunofluorescence and fluorescent histochemistry with CGRP and phalloidin, respectively. In addition, the origin of these nerve fibers was retrograde traced with a fluorescent neural tracer.

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Cancer Research

Two Flow Cytometric Approaches of NKG2D Ligand Surface Detection to Distinguish Stem Cells from Bulk Subpopulations in Acute Myeloid Leukemia
Henrik Landerer *1, Marlon Arnone *1, Ronja Wieboldt 2, Elsa Goersch 1, Anna M. Paczulla Stanger 3, Martina Konantz 1, Claudia Lengerke 1,2,3
1Department of Biomedicine, University Hospital Basel and University of Basel, 2Division for Hematology, University Hospital Basel and University of Basel, 3Department of Internal Medicine, Hematology and Oncology, University Hospital Tübingen

We present two different staining protocols for NKG2D ligand (NKG2DL) detection in human primary acute myeloid leukemia (AML) samples. The first approach is based on a fusion protein, able to recognize all known and potentially yet unknown ligands, while the second protocol relies on the addition of multiple anti-NKG2DL antibodies.

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Biology

Optical Sectioning and Visualization of the Intervertebral Disc from Embryonic Development to Degeneration
Florian Christof Bonnaire 1,2, Martina Feierabend 3, Julius Michael Wolfgart 1,4, Wolfram Breuer 5, Christian Walter 2, Ulf Krister Hofmann 1,2, Marina Danalache 1
1Laboratory of Cell Biology, Department of Orthopedic Surgery, University Hospital of Tübingen, 2Department of Orthopedic Surgery, University Hospital of Tübingen, 3Institute for Bioinformatics and Medical Informatics, Faculty of science of the University of Tübingen, 4Medical faculty of the University of Tübingen, 5Bavarian Health and Food Safety Authority

We present a method to investigate spatial chondrocyte organization in the anulus fibrosus of the intervertebral disc using an optical sectioning method.

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Bioengineering

Injection of Porcine Adipose Tissue-Derived Stroma Cells via Waterjet Technology
Jasmin Knoll *1, Marina Danalache *2, Walter Linzenbold 3, Markus Enderle 3, Tanja Abruzzese 1, Arnulf Stenzl 1, Wilhelm K. Aicher 1
1Department of Urology, University Hospital Tübingen, 2Department of Orthopaedic Surgery, University Hospital Tübingen, 3ERBE Elektromedizin GmbH Tübingen

We present a method of cell injection via needle free waterjet technology coupled with a sequela of post-delivery investigations in terms of cellular viability, proliferation, and elasticity measurements.

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Medicine

A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro
Wanjing Cen 1, Siegmar Reinert 1, Meltem Avci-Adali 2, Dorothea Alexander 1, Felix Umrath 1,3
1Department of Oral and Maxillofacial Surgery, University Hospital Tübingen, 2Department of Thoracic and Cardiovascular Surgery, University Hospital Tübingen, 3Department of Orthopaedic Surgery, University Hospital Tübingen

Here, we present a simple and effective assay procedure for resorption pit assays using calcium phosphate coated cell culture plates.

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Immunology and Infection

Polarization and Characterization of M1 and M2 Human Monocyte-Derived Macrophages on Implant Surfaces
Leila Mohammadnejad 1, Hannah Conrady 1, Madeline Mangold 1, Evi Kimmerle-Mueller 1, Annika Hechler 2, Christiane von Ohle 2, Frank Rupp 1, Stefanie Krajewski 1
1Department of Medical Materials Science & Technology, Institute for Biomedical Engineering, University Hospital Tübingen, 2Department of Conservative Dentistry and Periodontology, University Hospital Tübingen

Here, we present a detailed protocol for assessing the immunomodulatory potential of implant surfaces in vitro, aiming to improve the reliability and reproducibility of current protocols and promoting further research. Secretory cytokine profiles, mRNA expression, and cell surface markers were monitored using blood monocyte-derived macrophages to investigate macrophage polarization cultivated on titanium.

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