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Vrije Universiteit Amsterdam

10 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Depletion and Reconstitution of Macrophages in Mice
Shelley B. Weisser 1, Nico van Rooijen 2, Laura M. Sly 3
1Department of Graduate Studies, University of British Columbia , 2Department of Molecular Biology, Vrije Universiteit Amsterdam, 3Department of Pediatrics, University of British Columbia

Macrophages play a central role in homeostasis and pathology in many tissues. The protocol presented here describes methods for depleting macrophages in vivo, deriving polarized macrophages from bone marrow aspirates, and adoptively transferring macrophages into mice. These techniques allow determination of the role that polarized macrophages play in health and disease.

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Biology

Visualizing Cytoplasmic Flow During Single-cell Wound Healing in Stentor coeruleus
Mark Slabodnick 1,2, Bram Prevo 1,3, Peter Gross 1,4, Janet Sheung 1,5, Wallace Marshall 1,2
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.

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Developmental Biology

Streamlined 3D Cerebellar Differentiation Protocol with Optional 2D Modification
Dwayne B. Holmes 1, Vivi M. Heine 1,2
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.

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Behavior

Investigating the Deployment of Visual Attention Before Accurate and Averaging Saccades via Eye Tracking and Assessment of Visual Sensitivity
Luca Wollenberg 1,2, Heiner Deubel 1, Martin Szinte 1,3
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.

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JoVE Journal

Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree Parasponia andersonii
Titis A.K. Wardhani 1,2, Yuda Purwana Roswanjaya 1,2, Simon Dupin 1,3, Huchen Li 1,4, Sidney Linders 1, Marijke Hartog 1, Rene Geurts 1, Arjan van Zeijl 1
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.

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Medicine

A Postoperative Evaluation Guideline for Computer-Assisted Reconstruction of the Mandible
Gustaaf J. C. van Baar 1, Niels P. T. J. Liberton 2, Henri A. H. Winters 3, Lars Leeuwrik 1, Tymour Forouzanfar 1, Frank K. J. Leusink 1
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.

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JoVE Core

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. Brink 1, Cornelis J. de Ruiter 2, Geert J.P. Savelsbergh 2, Annemarijn Steijlen 3, Kaspar M.B. Jansen 3, Frans C.T. van der Helm 4, Edwin A. Goedhart 5, Doris van der Laan 6, Riemer J.K. Vegter 1, Koen A.P.M. Lemmink 1
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.

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Bioengineering

Quantification of Mouse Heart Left Ventricular Function, Myocardial Strain, and Hemodynamic Forces by Cardiovascular Magnetic Resonance Imaging
Mariah R. R. Daal 1, Gustav J. Strijkers 1,2, Claudia Calcagno 2, Ruslan R. Garipov 3, Rob C. I. Wüst 1,4, David Hautemann 5, Bram F. Coolen 1
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.

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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 Yeung 1,2
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.

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Neuroscience

Production of High-Yield Adeno Associated Vector Batches Using HEK293 Suspension Cells
Kimberly L. Pietersz 1, Paul J.H. Nijhuis 1, Matthijs H.M. Klunder 1, Joëlle van den Herik 1, Barbara Hobo 1, Fred de Winter 1, Joost Verhaagen 1,2
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.

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