Vrije Universiteit Amsterdam 12 articles published in JoVE Neuroscience Production of High-Yield Adeno Associated Vector Batches Using HEK293 Suspension Cells Kimberly L. Pietersz1, Paul J.H. Nijhuis1, Matthijs H.M. Klunder1, Joëlle van den Herik1, Barbara Hobo1, Fred de Winter1, Joost Verhaagen1,2 1Laboratory for Regeneration of Sensorimotor Systems, Netherlands Institute for Neuroscience, Royal Netherlands Academy of Arts and Sciences (KNAW), 2Centre for Neurogenomics and Cognitive Research, Amsterdam Neuroscience, Vrije Universiteit Amsterdam Here, a suspension HEK293 cell-based AAV production protocol is presented, resulting in reduced time and labor needed for vector production using components that are available for research purposes from commercial vendors. Medicine Robotic D3 Partial Duodenal Resection with Primary Side-to-Side Anastomosis Roberto Maria Montorsi1,2,3, Sofia Xenaki1,2,4, Sebastiaan Festen5, Paul Fockens2,6, Barbara A. J. Bastiaansen2,6, Freek Daams2,7, Olivier R. Busch1,2, M. G. Besselink1,2, HPB-Amsterdam, 1Amsterdam UMC, Department of Surgery, University of Amsterdam, 2Cancer Center Amsterdam, 3Department of General and Pancreatic Surgery, The Pancreas Institute, University of Verona Hospital Trust, 4Department of General Surgery, University Hospital of Heraklion Crete, 5Department of Surgery, OLVG, 6Amsterdam UMC, Department of Gastroenterology, University of Amsterdam, 7Amsterdam UMC, Department of Surgery, Vrije Universiteit This protocol presents a case of a robotic partial duodenal resection with primary side-to-side duodeno-jejunal reconstruction in a patient with a 5 cm duodenal stenosis. This is done at the third duodenal segment (D3) after an endoscopic mucosal resection (EMR) for a duodenal polyp. Biology Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro Natalija Bogunovic*1,2,3, Karlijn B. Rombouts*1,2, Kak Khee Yeung1,2 1Department of Vascular Surgery, Amsterdam UMC, Vrije Universiteit Amsterdam, 2Department of Physiology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, Vrije Universiteit Amsterdam, 3Laboratory of Experimental Cardiology, Department of Cardiology, Leiden University Medical Center This paper describes an explant culture-based method for the isolation and culturing of primary, patient-specific human aortic smooth muscle cells and dermal fibroblasts. Furthermore, a novel method is presented for measuring cell contraction and subsequent analysis, which can be used to study patient-specific differences in these cells. Medicine Robotic Central Pancreatectomy with Roux-en-Y Pancreaticojejunostomy Tess M.E. van Ramshorst1, Maurice J.W. Zwart1, Rogier P. Voermans2, Sebastiaan Festen3, Freek Daams4, Olivier R. Busch1, Matthijs W.N. Oomen5, Marc G. Besselink1, HPB-Amsterdam, 1Department of Surgery, Amsterdam UMC, University of Amsterdam, Cancer Center Amsterdam, 2Department of Gastroenterology, Amsterdam UMC, University of Amsterdam, Amsterdam Gastroenterology Endocrinology Metabolism, 3Department of Surgery, OLVG, 4Department of Surgery, Amsterdam UMC, Vrije Universiteit, 5 Robotic central pancreatectomy may be used in selected patients in experienced centers. This protocol presents all steps and the feasibility of a robotic central pancreatectomy with Roux-en-Y pancreaticojejunostomy in a 16-year-old adolescent patient. Bioengineering Quantification of Mouse Heart Left Ventricular Function, Myocardial Strain, and Hemodynamic Forces by Cardiovascular Magnetic Resonance Imaging Mariah R. R. Daal1, Gustav J. Strijkers1,2, Claudia Calcagno2, Ruslan R. Garipov3, Rob C. I. Wüst1,4, David Hautemann5, Bram F. Coolen1 1Department of Biomedical Engineering & Physics, Amsterdam University Medical Centers, Amsterdam Cardiovascular Sciences, University of Amsterdam, 2BioMedical Engineering and Imaging Institute, Icahn School of Medicine at Mount Sinai, 3MR Solutions Ltd., 4Laboratory for Myology, Department of Human Movement Sciences, Faculty of Behavioral and Movement Sciences, Vrije Universiteit Amsterdam, 5Medis medical imaging systems B.V. This study describes a comprehensive cardiovascular magnetic resonance imaging (CMR) protocol to quantify the left ventricular functional parameters of the mouse heart. The protocol describes the acquisition, post-processing, and analysis of the CMR images as well as assessment of different cardiac functional parameters. Behavior An Inertial Measurement Unit Based Method to Estimate Hip and Knee Joint Kinematics in Team Sport Athletes on the Field Bram J.C. Bastiaansen*1, Erik Wilmes*2, Michel S. Brink1, Cornelis J. de Ruiter2, Geert J.P. Savelsbergh2, Annemarijn Steijlen3, Kaspar M.B. Jansen3, Frans C.T. van der Helm4, Edwin A. Goedhart5, Doris van der Laan6, Riemer J.K. Vegter1, Koen A.P.M. Lemmink1 1Center for Human Movement Sciences, University Medical Center, University of Groningen, 2Department of Human Movement Sciences, Faculty of Behavioural and Movement Sciences, Vrije Universiteit Amsterdam, 3Emerging Materials, Department of Design Engineering, Delft University of Technology, 4Department of Biomechanical Engineering, Delft University of Technology, 5FIFA Medical Center, Royal Netherlands Football Association, 6Royal Dutch Hockey Association Monitoring athletes is essential for improving performance and reducing injury risk in team sports. Current methods to monitor athletes do not include the lower extremities. Attaching multiple inertial measurement units to the lower extremities could improve monitoring athletes in the field. Medicine A Postoperative Evaluation Guideline for Computer-Assisted Reconstruction of the Mandible Gustaaf J. C. van Baar1, Niels P. T. J. Liberton2, Henri A. H. Winters3, Lars Leeuwrik1, Tymour Forouzanfar1, Frank K. J. Leusink1 1Department of Oral and Maxillofacial Surgery/Pathology, Amsterdam UMC and Academic Centre for Dentistry Amsterdam (ACTA), Vrije Universiteit Amsterdam, 2Medical Technology, 3D Innovation Lab, Amsterdam UMC, Vrije Universiteit Amsterdam, 3Department of Plastic, Reconstructive and Hand Surgery, Amsterdam UMC, Vrije Universiteit Amsterdam Here, we propose a practical, feasible and reproducible evaluation guideline for computer-assisted reconstruction of the mandible in order to create uniformity between studies regarding postoperative accuracy evaluation. This protocol continues and specifies an earlier publication of this evaluation guideline. Genetics Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree Parasponia andersonii Titis A.K. Wardhani1,2, Yuda Purwana Roswanjaya1,2, Simon Dupin1,3, Huchen Li1,4, Sidney Linders1, Marijke Hartog1, Rene Geurts1, Arjan van Zeijl1 1Laboratory of Molecular Biology, Department of Plant Sciences, Wageningen University & Research, 2Center of Technology for Agricultural Production, Agency for the Assessment and Application of Technology (BPPT), 3Department of Ecological Science, Faculty of Earth and Life Sciences, Vrije Universiteit Amsterdam, 4Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, Beijing University of Agriculture Parasponia andersonii is a fast-growing tropical tree that belongs to the Cannabis family (Cannabaceae) and can form nitrogen-fixing root nodules in association with the rhizobium. Here, we describe a detailed protocol for reverse genetic analyses in P. andersonii based on Agrobacterium tumefaciens-mediated stable transformation and CRISPR/Cas9-based genome editing. Behavior Investigating the Deployment of Visual Attention Before Accurate and Averaging Saccades via Eye Tracking and Assessment of Visual Sensitivity Luca Wollenberg1,2, Heiner Deubel1, Martin Szinte1,3 1Allgemeine und Experimentelle Psychologie, Ludwig-Maximilians-Universität München, 2Graduate School of Systemic Neurosciences, Ludwig-Maximilians-Universität München, 3Department of Cognitive Psychology, Vrije Universiteit Amsterdam This experimental protocol combined eye tracking and the assessment of presaccadic visual sensitivity in a dual task paradigm, consisting of a free choice saccade task and a visual discrimination task, to investigate the deployment of visual spatial attention before both, accurate and averaging saccades. Developmental Biology Streamlined 3D Cerebellar Differentiation Protocol with Optional 2D Modification Dwayne B. Holmes1, Vivi M. Heine1,2 1Department of Pediatrics/Child Neurology, Amsterdam Neuroscience, VU University Medical Center, 2Department of Complex Trait Genetics, Center for Neurogenomics and Cognitive Research, Amsterdam Neuroscience, Vrije Universiteit Amsterdam We describe a simplified 3D differentiation protocol for hPSCs, using defined medium and reduced growth factors, capable of generating cell aggregates with early neuroepithelial structures and positive for cerebellar-associated markers, as well as an optional 2D modification for differentiating cells as a monolayer to generate functional neurons. Biology Novel Production Protocol for Small-scale Manufacture of Probiotic Fermented Foods Nieke Westerik1,2, Alex Paul Wacoo1,2,3, Wilbert Sybesma1, Remco Kort1,2,4,5 1Yoba for Life Foundation, 2Department of Molecular Cell Biology, Vrije Universiteit Amsterdam (VUA), 3Uganda Industrial Research Institute (UIRI), 4Micropia, Natura Artis Magistra, 5Department of Microbiology and Systems Biology, Netherlands Organisation for Applied Scientific Research (TNO) A production protocol is described for the small-scale production of probiotic fermented dairy drink with the aid of a novel bacterial starter culture. Biology Visualizing Cytoplasmic Flow During Single-cell Wound Healing in Stentor coeruleus Mark Slabodnick1,2, Bram Prevo1,3, Peter Gross1,4, Janet Sheung1,5, Wallace Marshall1,2 1Physiology Course, Marine Biological Laboratory, 2Department of Biochemistry & Biophysics, University of California San Francisco, 3Department of Physics and Astronomy, Vrije Universiteit Amsterdam, 4Max Planck Institute of Molecular Cell Biology and Genetics, 5Department of Physics, University of Illinois Urbana-Champaign The giant ciliate Stentor coeruleus is a classical system for studying regeneration and wound healing in single cells. By imaging Stentor cells simultaneously at low and high magnification it is possible to measure cytoplasmic flows before, during, and after wounding.
