University of Western Australia View Institution's Website 17 articles published in JoVE Bioengineering Decellularized Apple-Derived Scaffolds for Bone Tissue Engineering In Vitro and In Vivo Maxime Leblanc Latour1, Maryam Tarar2, Ryan J. Hickey1, Charles M. Cuerrier1, Isabelle Catelas3,4,5, Andrew E. Pelling1,2,6,7 1Department of Physics, University of Ottawa, 2Department of Biology, University of Ottawa, 3Department of Mechanical Engineering, University of Ottawa, 4Department of Surgery, University of Ottawa, 5Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 6Institute for Science, Society and Policy, University of Ottawa, 7SymbioticA, School of Human Sciences, University of Western Australia In this study, we detail methods of decellularization, physical characterization, imaging, and in vivo implantation of plant-based biomaterials, as well as methods for cell seeding and differentiation in the scaffolds. The described methods allow the evaluation of plant-based biomaterials for bone tissue engineering applications. Immunology and Infection Quantitative Polymerase Chain Reaction (qPCR)-Based Rapid Diagnosis of Helicobacter pylori Infection and Antibiotic Resistance Liang Wang*1,2,3, Jin-Xin Lai*1, Yu-Ting Si*1,4, Xu-Xia Cui1,4, Zeeshan Umar1,5, Xiao-Jun Ru1, Xin-Yu Zhang1, Zheng-Kang Li1, Alfred Chin Yen Tay5,6,7,8, Barry J. Marshall5,6,7,8, Guang-Hua Li1, Bing Gu1 1Laboratory Medicine, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, 2Division of Microbiology and Immunology, School of Biomedical Sciences, The University of Western Australia, 3Center for Precision Health, School of Medical and Health Sciences, Edith Cowan University, 4Medical Technology School of Xuzhou Medical University, 5Marshall Laboratory of Biomedical Engineering, School of Medicine, Shenzhen University, 6The Marshall Centre for Infectious Diseases Research and Training, The University of Western Australia, 7Marshall International Digestive Diseases Hospital, Zhengzhou University, 8Marshall Medical Research Center, Fifth Affiliated Hospital of Zhengzhou University The protocol presents a noninvasive method for the rapid diagnosis of Helicobacter pylori stomach infections through the string test and determines its antibiotic resistance to clarithromycin and levofloxacin using quantitative polymerase chain reaction (qPCR). Biology Fluorescence Lifetime Macro Imager for Biomedical Applications Rajannya Sen1, Alexander V. Zhdanov1, Ciaran Devoy2, Mark Tangney2, Liisa M. Hirvonen3, Andrei Nomerotski4, Dmitri B. Papkovsky1 1School of Biochemistry and Cell Biology, University College Cork, 2Cancer Research@UCC, University College Cork, 3Centre for Microscopy, Characterisation and Analysis (CMCA), The University of Western Australia, 4Physics Department, Brookhaven National Laboratory This paper describes the use of a new, fast optical imager for the macroscopic photoluminescence lifetime imaging of long decay emitting samples. The integration, image acquisition, and analysis procedures are described, along with the preparation and characterization of the sensor materials for the imaging and the application of the imager in studying biological samples. Immunology and Infection A Modified Murine Heterotopic Heart Transplant Protocol Matching Contemporary Standards of Aseptic Technique, Anesthesia, and Analgesia David A. Singer1, Gabrielle C. Musk2, Wen Hua Huang1,3, Liu Liu1, Jaskirat Kaur1, Monalyssa Watson1, Amy Prosser1, Michaela Lucas1,4, Andrew Lucas5 1Medical School, The University of Western Australia, 2Animal Care Services, The University of Western Australia, 3Western Australian Liver and Kidney Transplant Service, Sir Charles Gairdner Hospital, 4Department of Immunology, Sir Charles Gairdner Hospital and Pathwest Laboratory Medicine, 5School of Biomedical Sciences, The University of Western Australia The present paper describes a modified technique for heterotopic vascularized cardiac transplantation with updated aseptic technique, analgesia, and anesthesia. Behavior Controlled Rotation of Human Observers in a Virtual Reality Environment Michael Falconbridge1, Phillip Falconbridge2, David R. Badcock1 1School of Psychology, University of Western Australia, 2Falco Enterprise The controlled physical rotation of a human observer is desirable for certain experimental, recreational, and educational applications. This paper outlines a method for converting an office swivel chair into a medium for controlled physical rotation in a virtual reality environment. Medicine Applying a Three-dimensional Uniaxial Mechanical Stimulation Bioreactor System to Induce Tenogenic Differentiation of Tendon-Derived Stem Cells Ziming Chen*1, Peilin Chen*1, Rui Ruan*1, Lianzhi Chen1, Jun Yuan1, David Wood1, Tao Wang1, Ming Hao Zheng1 1Centre of Orthopaedic Translational Research, Medical School, University of Western Australia A three-dimensional uniaxial mechanical stimulation bioreactor system is an ideal bioreactor for tenogenic-specific differentiation of tendon-derived stem cells and neo-tendon formation. Medicine A Mouse Model of Incompletely Resected Soft Tissue Sarcoma for Testing (Neo)adjuvant Therapies Francois X. Rwandamuriye1,2, Breana J. Weston2,3, Terrance G. Johns2, W. Joost Lesterhuis1,2, Rachael M. Zemek2 1School of Biomedical Sciences, University of Western Australia, 2Telethon Kids Institute, University of Western Australia, 3College of Science, Health, Engineering and Education, Murdoch University In this protocol, we describe a mouse model of incomplete surgical resection of soft tissue sarcoma for testing (neo)adjuvant therapies. Medicine Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology Michael A. Pearen*1, Hong Kiat Lim*1, Francis D. Gratte2,3, Manuel A. Fernandez-Rojo1,4,5, Sujeevi K. Nawaratna6, Geoffrey N. Gobert7, John K. Olynyk8,9, Janina E. E. Tirnitz-Parker2,10, Grant A. Ramm1,4 1Hepatic Fibrosis Group, QIMR Berghofer Medical Research Institute, 2School of Pharmacy and Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, 3School of Veterinary and Life Sciences, Murdoch University, 4School of Medicine, The University of Queensland, 5Madrid Institute for Advanced Studies (IMDEA) in Food, CEI UAM+CSIC, 6School of Medicine, Griffith University, 7 This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology. Bioengineering Growth of Human and Sheep Corneal Endothelial Cell Layers on Biomaterial Membranes Jennifer Walshe1, Najla Al Khaled Abdulsalam2, Shuko Suzuki1, Traian V. Chirila1,3,4,5,6,7,8, Damien G. Harkin1,3,4 1Queensland Eye Institute, 2King Faisal University, 3School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, 4Institute of Health and Biomedical Innovation, Queensland University of Technology, 5Science and Engineering Faculty, Queensland University of Technology, 6Faculty of Medicine, University of Queensland, 7Australian Institute for Bioengineering and Nanotechnology, University of Queensland, 8Faculty of Science, University of Western Australia This protocol describes the critical steps required to establish and grow corneal endothelial cell cultures from explants of human or sheep tissue. A method for subculturing corneal endothelial cells on membranous biomaterials is also presented. Genetics Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry Sarah E. Ashley1,2, Braydon A. Meyer2,3, Justine A. Ellis2,3,4, David J. Martino2,3,5 1Molecular Genetics of Chronic Inflammation and Allergic Disease, Max-Delbrück Center for Molecular Medicine, 2 Identification of genetic variants contributing to complex human disease allows us to identify novel mechanisms. Here, we demonstrate a multiplex genotyping approach to candidate genes or gene pathway analysis that maximizes the coverage at low cost and is amenable to cohort-based studies. Genetics A Fast Silver Staining Protocol Enabling Simple and Efficient Detection of SSR Markers using a Non-denaturing Polyacrylamide Gel Ling Huang*1, Xiaohui Deng*1, Ronghua Li1, Yanshi Xia1, Guihua Bai2, Kadambot H.M. Siddique3, Peiguo Guo1 1College of Life Sciences, Guangzhou University, 2Hard Winter Wheat Genetics Research Unit, United States Department of Agriculture - Agricultural Research Service, 3The UWA Institute of Agriculture, University of Western Australia Here, we report a simple and low-cost silver staining protocol which requires only three reagents and 7 min of processing, and is suitable for fast generation of high-quality SSR data in the genetic analysis. Biochemistry Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana Wenhui Lyu1, Jennifer Selinski1, Lu Li1, David A. Day2, Monika W. Murcha3, James Whelan1, Yan Wang1 1ARC Centre of Excellence in Plant Energy Biology, Department of Animal, Plant and Soil Science, School of Life Science, La Trobe University, 2School of Biological Sciences, Flinders University, 3ARC Centre of Excellence in Plant Energy Biology, University of Western Australia As mitochondria are only a small percentage of the plant cell, they need to be purified for a range of studies. Mitochondria can be isolated from a variety of plant organs by homogenization, followed by differential and density gradient centrifugation to obtain a highly purified mitochondrial fraction. Biology The Murine Choline-Deficient, Ethionine-Supplemented (CDE) Diet Model of Chronic Liver Injury Jully Gogoi-Tiwari1, Julia Köhn-Gaone1, Corey Giles2, Dirk Schmidt-Arras3, Francis D. Gratte1,4, Caryn L. Elsegood1, Geoffrey W. McCaughan5,6,7, Grant A. Ramm8,9, John K. Olynyk10,11, Janina E.E. Tirnitz-Parker1,12 1School of Biomedical Sciences & Curtin Health Innovation Research Institute, Curtin University, 2School of Public Health & Curtin Health Innovation Research Institute, Curtin University, 3Institute of Biochemistry, Christian-Albrechts-University, 4School of Veterinary and Life Sciences, Murdoch University, 5Centenary Institute of Cancer Medicine and Cell Biology, The University of Sydney, 6Royal Prince Alfred Hospital, 7A.W. Morrow Gastroenterology and Liver Centre, 8QIMR Berghofer Medical Research Institute, 9Faculty of Medicine and Biomedical Sciences, The University of Queensland, 10Fiona Stanley and Fremantle Hospitals, 11School of Medical and Health Sciences, Edith Cowan University, 12School of Medicine and Pharmacology, University of Western Australia Here we describe a common method to induce chronic liver injury in mice by feeding of a choline-deficient and ethionine-supplemented (CDE) diet. We demonstrate health monitoring, liver perfusion, isolation, and preservation. A time course of six weeks can inform about liver injury, pathohistology, fibrosis, inflammatory, and liver progenitor cell responses. Biology Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins Kathryn A. Ramsey1,2, Zachary L. Rushton1, Camille Ehre1,3 1Marsico Lung Institute/CF Center, University of North Carolina at Chapel Hill, 2Telethon Kids Institute, University of Western Australia, 3Department of Pediatrics, University of North Carolina at Chapel Hill Mucins are high-molecular-weight glycoconjugates, with size ranging from 0.2 to 200 megadalton (MDa). As a result of their size, mucins do not penetrate conventional polyacrylamide gels and require larger pores for separation. We provide a detailed protocol for mucin agarose gel electrophoresis to assess relative quantification and study polymer assembly. Biology Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro Marcus K. Giacci1, Nathan S. Hart2, Richard V. Hartz1, Alan R. Harvey3, Stuart I. Hodgetts3, Melinda Fitzgerald1 1Experimental and Regenerative Neurosciences, School of Animal Biology, The University of Western Australia, 2School of Animal Biology and The Oceans Institute, The University of Western Australia, 3Experimental and Regenerative Neurosciences, School of Anatomy, Physiology and Human Biology, The University of Western Australia Non-coherent Xenon light was passed through narrow-band interference and neutral density filters to deliver light of varying wavelength and intensity to cultured cells. This protocol was used to assess the effects of red/near-infrared light therapy on production of reactive species in vitro: no effects were observed using the tested parameters. Behavior Measuring Sensitivity to Viewpoint Change with and without Stereoscopic Cues Jason Bell1,2, Edwin Dickinson2, David R. Badcock2, Frederick A. A. Kingdom3 1Research School of Psychology, Australian National University, 2School of Psychology, University of Western Australia, 3McGill Vision Research, Department of Ophthalmology, McGill University We discuss a novel method forviewpoint-rotation of visual stimuli, and demonstrate using a mirror stereoscopethe three-dimensional percept of rotation-in-depth. The technique can be used to investigate the role of stereoscopic cues in encoding viewpoint-rotated figures. Biology Regular Care and Maintenance of a Zebrafish (Danio rerio) Laboratory: An Introduction Avdesh Avdesh*1,2, Mengqi Chen*1,3, Mathew T. Martin-Iverson1,2,4, Alinda Mondal1,3, Daniel Ong1, Stephanie Rainey-Smith1,3, Kevin Taddei1,3, Michael Lardelli5, David M. Groth6, Giuseppe Verdile1,3, Ralph N. Martins1,2,3,7 1Centre of Excellence for Alzheimer's Disease Research and Care, School of Medical sciences, Edith Cowan University, 2Centre for Clinical Research in Neuropsychiatry, Graylands Hospital, University of Western Australia, 3McCusker Alzheimer's Research foundation, 4School of Medicine and Pharmacology, University of Western Australia, 5Department of Molecular and Biomedical Sciences, University of Adelaide, 6School of Biomedical Sciences, Curtin University of Technology, 7School of Psychiatry and Clinical Neurosciences, University of Western Australia This protocol outlines regular maintenance and care to maintain optimal conditions for zebrafish husbandry. The video illustrates the protocol for system maintenance, regular housing, feeding, breeding, and raising of zebrafish larvae.
