university hospital basel

9 ARTICLES PUBLISHED IN JoVE

Medicine

Evaluation of Stem Cell Properties in Human Ovarian Carcinoma Cells Using Multi and Single Cell-based Spheres Assays

Hui Wang^1,2, Anna Paczulla¹, 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.

Chemistry

An ELISA Based Binding and Competition Method to Rapidly Determine Ligand-receptor Interactions

Mohameedyaseen Syedbasha¹, Janina Linnik^1,2,3, Deanna Santer⁴, Daire O'Shea⁵, Khaled Barakat^4,6, Michael Joyce⁴, Nina Khanna⁷, D. Lorne Tyrrell⁴, Michael Houghton⁴, Adrian Egli^1,8

¹Applied Microbiology Research, Department of Biomedicine, University of Basel, ²Department of Biosystems Science and Engineering, ETH Zurich, and Swiss Institute of Bioinformatics, ³Swiss Institute of Bioinformatics, ⁴Li Ka Shing Institute for Virology, University of Alberta, ⁵Regional Infectious Diseases Unit, University of Edinburgh, ⁶Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, ⁷Infection Biology, Department of Biomedicine, University of Basel, ⁸Clinical Microbiology, University Hospital Basel

The presented protocols describe two enzyme-linked immunosorbent assay (ELISA) based techniques for the rapid investigation of ligand-receptor interactions: The first assay allows the determination of dissociation constant between ligand and receptor. The second assay enables a rapid screening of blocking peptides for ligand-receptor interactions.

Developmental Biology

An Enzyme- and Serum-free Neural Stem Cell Culture Model for EMT Investigation Suited for Drug Discovery

Martin H. M. Sailer¹, Durga Sarvepalli², Catherine Brégère³, Urs Fisch³, Marin Guentchev⁴, Michael Weller⁶, Raphael Guzman³, Bernhard Bettler¹, Arkasubhra Ghosh^*2, Gregor Hutter^*5

¹Dept. of Biomedicine, Pharmacenter, University of Basel, ²Molecular Signalling and Gene Therapy, Narayana Nethralaya Foundation, Narayana Health City, ³Brain Ischemia and Regeneration, Department of Biomedicine, University Hospital Basel, ⁴Department of Neurosurgery, Klinikum Idar-Oberstein, ⁵Department of Neurosurgery and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, ⁶Department of Neurology, Laboratory of Molecular Neuro Oncology, University Hospital of Zurich

Epithelial to mesenchymal transition (EMT) allows cancers to become invasive. To investigate EMT, a neural stem cell (NSC)-based in vitro model devoid of serum and enzymes is described. This standardized system allows quantitative and qualitative assessment of cell migration, gene and protein expression. The model is suited for drug discovery.

Immunology and Infection

Development of an Antigen-driven Colitis Model to Study Presentation of Antigens by Antigen Presenting Cells to T Cells

Valerio Rossini¹, Katarina Radulovic², Christian U. Riedel³, Jan Hendrik Niess²

¹APC Microbiome Institute, University College Cork, ²Division of Gastroenterology and Hepatology, University Hospital Basel, ³Institute of Microbiology and Biotechnology, University of Ulm

In this antigen-driven colitis model, OT-II CD4⁺ T cells expressing a red fluorescent protein were adoptively transferred into RAG^-/- mice that express a green fluorescent protein in mononuclear phagocytes (MPs). The hosts were challenged with Escherichia coli (E.coli) expressing the ovalbumin protein (OVA) fused to a cyan fluorescent protein (CFP).

Medicine

An Optimized Hemagglutination Inhibition (HI) Assay to Quantify Influenza-specific Antibody Titers

Lukas Kaufmann^*1, Mohammedyaseen Syedbasha^*1, Dominik Vogt¹, Yvonne Hollenstein¹, Julia Hartmann¹, Janina E. Linnik^1,2,3, Adrian Egli^1,4

¹Applied Microbiology Research, Department of Biomedicine, University of Basel, ²Department of Biosystems Science and Engineering, ETH Zurich, ³Swiss Institute of Bioinformatics, ⁴Clinical Microbiology, University Hospital Basel

The presented protocols describe how to perform a hemagglutination inhibition assay to quantify influenza-specific antibody titers from serum samples of influenza vaccine recipients. The first assay determines optimal viral antigen concentrations by hemagglutination. The second assay quantifies influenza-specific antibody titers by hemagglutination inhibition.

JoVE Journal

Injections of Lipopolysaccharide into Mice to Mimic Entrance of Microbial-derived Products After Intestinal Barrier Breach

Katarina Radulovic¹, Rachel Mak’Anyengo¹, Berna Kaya¹, Anna Steinert¹, Jan Hendrik Niess^1,2

¹Department of Biomedicine, University of Basel, ²Division of Gastroenterology and Hepatology, University Hospital of Basel

Here a protocol to mimic the entrance of bacterial-derived compounds after intestinal barrier breach is presented. A low sublethal dose of lipopolysaccharide was injected systemically into mice, which were monitored for 24 h post-injection. The expression of pro-inflammatory cytokines was determined at several time points in spleen, liver, and colon.

Developmental Biology

Induction of Endothelial Differentiation in Cardiac Progenitor Cells Under Low Serum Conditions

Michika Mochizuki^*1, Giacomo Della Verde^*1, Habiba Soliman¹, Otmar Pfister^1,2, Gabriela M. Kuster^1,2

¹Department of Biomedicine, University Hospital and University of Basel, ²Division of Cardiology, University Hospital Basel

This protocol describes an endothelial differentiation technique for cardiac progenitor cells. It particularly focuses on how serum concentration and cell-seeding density affect the endothelial differentiation potential.

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 Cui¹, Chen She¹, Hui Wang¹, Shuang Wu¹, Ling Zou¹, Jianliang Zhang¹, Wanzhu Bai¹

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.

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 Wieboldt², Elsa Goersch¹, Anna M. Paczulla Stanger³, Martina Konantz¹, 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.