University Hospital Tubingen 8 articles published in JoVE Bioengineering Addressing Practical Issues in Atomic Force Microscopy-Based Micro-Indentation on Human Articular Cartilage Explants Cyril Daniel1, Dorothea Alexander2, Felix Umrath1,2, Marina Danalache1 1Laboratory of Cell Biology, Department of Orthopaedic Surgery, University Hospital Tübingen, 2Department of Oral and Maxillofacial Surgery, University Hospital Tübingen We present a step-by-step approach to identify and address the most common problems associated with atomic force microscopy micro-indentations. We exemplify the emerging problems on native human articular cartilage explants characterized by various degrees of osteoarthritis-driven degeneration. Medicine A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro Wanjing Cen1, Siegmar Reinert1, Meltem Avci-Adali2, Dorothea Alexander1, Felix Umrath1,3 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. Bioengineering Injection of Porcine Adipose Tissue-Derived Stroma Cells via Waterjet Technology Jasmin Knoll*1, Marina Danalache*2, Walter Linzenbold3, Markus Enderle3, Tanja Abruzzese1, Arnulf Stenzl1, Wilhelm K. Aicher1 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. Biology Optical Sectioning and Visualization of the Intervertebral Disc from Embryonic Development to Degeneration Florian Christof Bonnaire1,2, Martina Feierabend3, Julius Michael Wolfgart1,4, Wolfram Breuer5, Christian Walter2, Ulf Krister Hofmann1,2, Marina Danalache1 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. 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 Lengerke1,2,3 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. Bioengineering Application of Atomic Force Microscopy to Detect Early Osteoarthritis Marina Danalache*1, Aadhya Tiwari*1, Viktor Sigwart1,2, Ulf Krister Hofmann1,3 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). 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 Friedrich1, Vivien Hanel1, Peter Kobelt1, Miriam Goebel-Stengel1,2,3, Matthias Rose1,4, Andreas Stengel1,3 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. Medicine Evaluation of Stem Cell Properties in Human Ovarian Carcinoma Cells Using Multi and Single Cell-based Spheres Assays Hui Wang1,2, Anna Paczulla1, Claudia Lengerke1,2 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.
