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VU University medical center

9 ARTICLES PUBLISHED IN JoVE

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Medicine

High Content Screening in Neurodegenerative Diseases
Shushant Jain 1, Ronald E. van Kesteren 2, Peter Heutink 1
1Department of Clinical Genetics, VU University Medical Center, 2Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam

We describe a methodology combining automated cell culturing with high-content imaging to visualize and quantify multiple cellular processes and structures, in a high-throughput manner. Such methods can aid in the further functional annotation of genomes as well as identify disease gene networks and potential drug targets.

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Immunology and Infection

Infection of Zebrafish Embryos with Intracellular Bacterial Pathogens
Erica L. Benard 1, Astrid M. van der Sar 2, Felix Ellett 3, Graham J. Lieschke 3, Herman P. Spaink 1, Annemarie H. Meijer 1
1Department of Molecular Cell Biology, Institute of Biology, Leiden University, 2Department of Medical Microbiology and Infection Control, VU University Medical Center, 3Australian Regenerative Medicine Institute, Monash University

Transparent zebrafish embryos have proved useful model hosts to visualize and functionally study interactions between innate immune cells and intracellular bacterial pathogens, such as Salmonella typhimurium and Mycobacterium marinum. Micro-injection of bacteria and multi-color fluorescence imaging are essential techniques involved in the application of zebrafish embryo infection models.

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Medicine

Measuring Cardiac Autonomic Nervous System (ANS) Activity in Children
Aimée E. van Dijk *1,2, René van Lien *3,4, Manon van Eijsden 2,5, Reinoud J. B. J. Gemke 6, Tanja G. M. Vrijkotte 1, Eco J. de Geus 3,4
1Department of Public Health, Academic Medical Center - University of Amsterdam, 2Department of Epidemiology, Documentation and Health Promotion, Public Health Service of Amsterdam (GGD), 3Department of Biological Psychology, VU University, 4EMGO+ Institute, VU University Medical Center, 5Institute of Health Sciences, VU University, 6Department of Pediatrics, VU University Medical Center

Measurement of autonomic nervous system activity usually confines the researcher and participant to the laboratory, which may provide an intimidating environment to children. The VU University Ambulatory Monitoring System (VU-AMS) device can record cardiac autonomic control in any setting. The VU-AMS proved very amenable to testing in children.

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Cancer Research

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models
Rocco Sciarrillo 1,2,3, Anna Wojtuszkiewicz 1, Irsan E. Kooi 4, Valentina E. Gómez 3, Ugo Boggi 5, Gerrit Jansen 6, Gert-Jan Kaspers 1,7, Jacqueline Cloos *1, Elisa Giovannetti *3,8,9
1Department of Pediatric Oncology/Hematology, VU University Medical Center, 2Department of Hematology, VU University Medical Center, 3Department of Medical Oncology, VU University Medical Center, 4Department of Clinical Genetics, VU University Medical Center, 5Division of General and Transplant Surgery, Azienda Ospedaliera Universitaria Pisana, Universita’ di Pisa, 6Amsterdam Immunology and Rheumatology Center, VU University Medical Center, 7Princess Máxima Center for Pediatric Oncology, 8Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, 9Institute of Nanoscience and Nanotechnology, CNR-Nano

Here we describe a protocol aimed at investigating the impact of aberrant splicing on drug resistance in solid tumors and hematological malignancies. To this goal, we analyzed the transcriptomic profiles of parental and resistant in vitro models through RNA-seq and established a qRT-PCR based method to validate candidate genes.

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Medicine

Combined Intravital Microscopy and Contrast-enhanced Ultrasonography of the Mouse Hindlimb to Study Insulin-induced Vasodilation and Muscle Perfusion
Alexander H. Turaihi 1, Erik M. van Poelgeest 1, Victor W. M. van Hinsbergh 1, Erik H. Serné 2, Yvo M. Smulders 2, Etto C. Eringa 1
1Laboratory for Physiology, Institute for Cardiovascular Research (ICaR-VU), VU University Medical Center, 2Department of Internal Medicine, Institute for Cardiovascular Research (ICaR-VU), VU University Medical Center

Insulin-induced vasodilation regulates muscle perfusion and increases the microvascular surface area (microvascular recruitment) available for solute exchange between blood and tissue interstitium. Combined intravital microscopy and contrast-enhanced ultrasonography is presented to simultaneously assess insulin's action at the larger vessels and the microcirculation in vivo.

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Cancer Research

Comprehensive Protocol to Sample and Process Bone Marrow for Measuring Measurable Residual Disease and Leukemic Stem Cells in Acute Myeloid Leukemia
Jacqueline Cloos *1,2, Jeffrey R. Harris *3, Jeroen J.W.M. Janssen 1, Angele Kelder 1, F. Huang 3, Gerrit Sijm 1, Maike Vonk 1, Alexander N. Snel 1, Jennifer R. Scheick 1, Willemijn J. Scholten 1, Jannemieke Carbaat-Ham 1, Dennis Veldhuizen 1, Diana Hanekamp 1, Yvonne J.M. Oussoren-Brockhoff 1, Gertjan J.L. Kaspers 2,4, Gerrit J. Schuurhuis 1, A. Kate Sasser 3, Gert Ossenkoppele 1
1Department of Hematology, VU University Medical Center, 2Pediatric Oncology/Hematology, VU University Medical Center, 3Janssen Research & Development, LLC, 4Princess Máxima Center for Pediatric Oncology

Detection of minimal or measurable residual disease (MRD) is an important prognostic biomarker for refining risk assessment and predicting relapse in acute myeloid leukemia (AML). These comprehensive guidelines and recommendations with best practices for consistent and accurate identification and detection of MRD, may aid in making effective AML treatment decisions.

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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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Immunology and Infection

In Vitro Microfluidic Disease Model to Study Whole Blood-Endothelial Interactions and Blood Clot Dynamics in Real-Time
Xue D. Manz 1, Hugo J. Albers 2,3, Petr Symersky 4, Jurjan Aman 1, Andries D. van der Meer 3, Harm Jan Bogaard 1, Robert Szulcek 1,5,6
1Department of Pulmonary Diseases, Amsterdam UMC, VU University Medical Center, Amsterdam Cardiovascular Sciences (ACS), 2BIOS Lab-on-a-Chip group, University of Twente, 3Applied Stem Cell Technologies Group, University of Twente, 4Department of Cardio-thoracic Surgery, Amsterdam UMC, VU University Medical Center, 5Institute of Physiology, Charité-Universitätsmedizin, 6German Heart Center

We present an in vitro vascular disease model to investigate whole blood interactions with patient-derived endothelium. This system allows the study of thrombogenic properties of primary endothelial cells under various circumstances. The method is especially suited to evaluate in situ thrombogenicity and anticoagulation therapy during different phases of coagulation.

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Biology

Automated Production of Human Induced Pluripotent Stem Cell-Derived Cortical and Dopaminergic Neurons with Integrated Live-Cell Monitoring
Ashutosh Dhingra *1, Joachim Täger *1, Elisangela Bressan *1, Salvador Rodriguez-Nieto 1, Manmeet-Sakshi Bedi 1, Stefanie Bröer 1, Eldem Sadikoglou 1, Noémia Fernandes 2, Melissa Castillo-Lizardo 1, Patrizia Rizzu 2, Peter Heutink 1,3
1Genome Biology of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), 2Applied Genomics for Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), 3Department for Neurodegenerative Diseases, Hertie Institute for Clinical Brain Research, University of Tübingen

We show the automation of human induced pluripotent stem cell (hiPSC) cultures and neuronal differentiations compatible with automated imaging and analysis.

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