Queen Mary University of London View Institution's Website 11 articles published in JoVE Biology A Triple Culture Cell System Modeling the Human Blood-Brain Barrier Eleonora Rizzi*1, Clémence Deligne*1, Lucie Dehouck1, Roberta Bilardo2, Yasutero Sano3, Fumitaka Shimizu3, Takashi Kanda3, Marina Resmini2, Fabien Gosselet1, Marie-Pierre Dehouck1, Caroline Mysiorek1 1Univ. Artois, UR 2465, Laboratoire de la Barrière Hémato-Encéphalique (LBHE), 2Queen Mary University of London, Department of Chemistry, 3Yamaguchi University Graduate School of Medicine, Department of Neurology and Clinical Neuroscience This protocol describes a method to establish a human blood-brain barrier (BBB) in vitro model. The endothelial cells and pericytes are seeded on each side of an insert filter (blood compartment), and astrocytes are seeded in the bottom well (brain compartment). The model characterized was used for nanoparticles transport experiments. Cancer Research Generation of Orthotopic Pancreatic Tumors and Ex vivo Characterization of Tumor-Infiltrating T Cell Cytotoxicity Sarah Spear1,2, Iain A McNeish1, Melania Capasso2,3 1Division of Cancer, Department of Surgery and Cancer, Imperial College London, 2Centre for Cancer and Inflammation, Barts Cancer Institute, Queen Mary University of London, 3German Center for Neurodegenerative Diseases (DZNE) This protocol describes the surgical generation of orthotopic pancreatic tumors and the rapid digestion of freshly isolated murine pancreatic tumors. Following digestion, viable immune cell populations can be used for further downstream analysis, including ex vivo stimulation of T cells for intracellular cytokine detection by flow cytometry. Biology Tissue Collection of Bats for -Omics Analyses and Primary Cell Culture Laurel R. Yohe*1,2, Paolo Devanna*3, Kalina T.J. Davies4, Joshua H.T. Potter4, Stephen J. Rossiter4, Emma C. Teeling5, Sonja C. Vernes*3,6, Liliana M. Dávalos*2,7 1Department of Geology & Geophysics, Yale University, 2Department of Ecology & Evolution, Stony Brook University, 3Neurogenetics of Vocal Communication, Max Planck Institute for Psycholinguistics, 4School of Biological and Chemical Sciences, Queen Mary University of London, 5School of Biology & Environmental Science, University College Dublin, 6Donders Institute for Brain, Cognition and Behavior, 7Consortium for Inter-Disciplinary Environmental Research, Stony Brook University This is a protocol for the optimal tissue preparation for genomic, transcriptomic, and proteomic analyses of bats caught in the wild. It includes protocols for bat capture and dissection, tissue preservation, and cell culturing of bat tissue. Engineering Design and Implementation of a Bespoke Robotic Manipulator for Extra-corporeal Ultrasound Shuangyi Wang1, James Housden1, Yohan Noh1, Anisha Singh2, Junghwan Back3, Lukas Lindenroth3, Hongbin Liu3, Joseph Hajnal1, Kaspar Althoefer4, Davinder Singh2, Kawal Rhode1 1 This paper introduces the design and implementation of a bespoke robotic manipulator for extra-corporeal ultrasound examination. The system has five degrees of freedom with lightweight joints made by 3D printing and a mechanical clutch for safety management. Bioengineering Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex Rong Li*1, Jinxin Miao*1,2, Zhiqiang Fan1, SeokHwan Song3, Il-keun Kong3,4, Yaohe Wang2,5, Zhongde Wang1 1Department of Animal, Dairy, and Veterinary Sciences, Utah State University, 2National Centre for International Research in Cell and Gene Therapy, Sino-British Research Centre, School of Basic Medical Sciences, Zhengzhou University, 3Department of Animal Science Division of Applied Life Science (BK21 Plus), Gyeongsang National University, 4Institute of Agriculture and Life Science, Gyeongsang National University, 5Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London Pronuclear (PN) injection of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein-9 nuclease (CRISPR/Cas9) system is a highly efficient method for producing genetically engineered golden Syrian hamsters. Herein, we describe the detailed PN injection protocol for the production of gene knockout hamsters with the CRISPR/Cas9 system. Immunology and Infection A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors Ming Yuan*1, Pengju Wang*2,3, Louisa S. Chard1, Nicholas R. Lemoine1,2,3, Yaohe Wang1,2,3 1Center for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, 2Sino-British Research Centre for Molecular Oncology, National Center for International Research in Cell and Gene Therapy, Zhengzhou University, 3School of Basic Medical Sciences, Academy of Medical Sciences, Zhengzhou University Vaccinia virus (VV) has been widely used in biomedical research and the improvement of human health. This article describes a simple, highly efficient method to edit the VV genome using a CRISPR-Cas9 system. Biology A Fast Air-dry Dropping Chromosome Preparation Method Suitable for FISH in Plants Lala Aliyeva-Schnorr1, Lu Ma1,2, Andreas Houben1 1Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), 2School of Biological and Chemical Sciences, Queen Mary University of London A protocol is described for the preparation of high-quality mitotic plant chromosome spreads by a fast air-dry dropping method suitable for the FISH detection of single and high copy DNA probes. Medicine Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies Enza Lonardo1,2, Michele Cioffi1, Patricia Sancho1,3, Shanthini Crusz3, Christopher Heeschen1,3 1Stem Cells & Cancer Group, Molecular Pathology Program, Spanish National Cancer Research Center, 2Institute for Research in Biomedicine (IRB Barcelona), 3Center for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided. Developmental Biology Dual Labeling of Neural Crest Cells and Blood Vessels Within Chicken Embryos Using ChickGFP Neural Tube Grafting and Carbocyanine Dye DiI Injection Jean-Marie Delalande1,2, Nikhil Thapar1, Alan J. Burns1,3 1Birth Defects Research Centre, UCL Institute of Child Health, 2Blizard Institute, Centre for Digestive Diseases, Queen Mary University of London, Barts and The London School of Medicine and Dentistry, 3Department of Clinical Genetics, Erasmus University Medical Center, Rotterdam Here we report dual labeling of neural crest cells and blood vessels using chickGFP neural tube intraspecies grafting combined with intra-vascular DiI injection. This experimental technique allows us to simultaneously visualize and study development of the NCC-derived (enteric) nervous system and the vascular system, during organogenesis. Medicine The Use of Reverse Phase Protein Arrays (RPPA) to Explore Protein Expression Variation within Individual Renal Cell Cancers Fiach C. O'Mahony1, Jyoti Nanda1, Alexander Laird1, Peter Mullen2, Helen Caldwell3, Ian M. Overton4, Lel Eory4, Marie O'Donnell1,5, Dana Faratian6, Thomas Powles7, David J. Harrison1,2, Grant D. Stewart1 1Edinburgh Urological Cancer Group, University of Edinburgh, 2School of Medicine, University of St Andrews, 3Division of Pathology, University of Edinburgh, 4MRC Human Genetics Unit, MRC IGMM, University of Edinburgh, 5Department of Pathology, Western General Hospital, 6Breakthrough Breast Cancer Research Unit, University of Edinburgh, 7St Bartholomew's Cancer Institute, Experimental Cancer Medicine Centre, Queen Mary University of London RPPA enables the protein expression of hundreds of samples, printed on nitrocellulose slides to be interrogated simultaneously, using fluorescently labelled antibodies. This technique has been applied to study the effect of drug treatment heterogeneity within clear cell renal carcinoma. Immunology and Infection Detection of Invasive Pulmonary Aspergillosis in Haematological Malignancy Patients by using Lateral-flow Technology Christopher Thornton1, Gemma Johnson2, Samir Agrawal3 1Biosciences, University of Exeter, 2BICMS, Queen Mary University of London, 3St. Bartholomew's Hospital and The London NHS Trust A rapid and accurate point-of-care test for invasive pulmonary aspergillosis is presented. It takes advantage of lateral-flow technology using a specific monoclonal antibody that binds to an Aspergillus antigen secreted during pulmonary infections. The assay is compatible with serum and brochoalveolar lavage and represents a novel adjunct test for disease diagnosis.