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Cardiff University

6 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models
Lee Parry 1, Madeleine Young 1, Fatima El Marjou 2, Alan Richard Clarke 1
1European Cancer Stem Cell Research Institute, Cardiff University, 2Institut Curie

Intestinal epithelial stem cells (ISCs) are intermingled with Paneth cells. These cells are differentiated progeny of the ISC, which support the ISCs and provide antibacterial protection. Here we demonstrate how we used transgenic conditional mouse models to establish that Paneth cells play a crucial role in maintaining the intestinal epithelia.

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Immunology and Infection

Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens
Bruce J. MacLachlan *1, Alexander Greenshields-Watson *1, Georgina H Mason *1, Andrea J Schauenburg 1, Valentina Bianchi 1,2,3, Pierre J Rizkallah 1, Andrew K Sewell 1, Anna Fuller 1, David K Cole 1
1Division of Infection and Immunity and Systems Immunity Research Institute, Cardiff University, 2Department of Oncology, University Hospital of Lausanne (CHUV), 3Ludwig Insitutue for Cancer Research, Lausanne Branch, University of Lausanne

Here, we describe methods that we commonly employ in the laboratory to determine how the nature of the interaction between the T-cell receptor and tumor antigens, presented by human leukocyte antigens, governs T-cell functionality; these methods include protein production, X-ray crystallography, biophysics, and functional T-cell experiments.

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Education

Method for Recording Broadband High Resolution Emission Spectra of Laboratory Lightning Arcs
D. Mitchard 1, D. Clark 1, D. Carr 1, C. Stone 1, A. Haddad 1
1Morgan-Botti Lightning Laboratory, Advanced High Voltage Research Centre, School of Engineering, Cardiff University

Emission spectroscopy techniques have traditionally been used to analyze inherently random lightning arcs occurring in nature. In this paper, a method developed to obtain the emission spectroscopy from reproducible lightning arcs generated within a laboratory environment is described.

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Neuroscience

In vitro Quantitative Imaging Assay for Phagocytosis of Dead Neuroblastoma Cells by iPSC-Macrophages
Hazel Hall-Roberts 1,2,3, Elena Di Daniel 2, William S. James 1, John B. Davis 2, Sally A. Cowley 1
1James Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford, 2Alzheimer’s Research UK Oxford Drug Discovery Institute, Nuffield Department of Medicine Research Building, University of Oxford, 3UK Dementia Research Institute, Cardiff University

Neurodegenerative diseases are associated with dysregulated microglia functions. This article outlines an in vitro assay of phagocytosis of neuroblastoma cells by iPSC-macrophages. Quantitative microscopy readouts are described for both live-cell time-lapse imaging and fixed-cell high-content imaging.

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Immunology and Infection

Lentiviral Vector Preparation for Efficient Gene and MicroRNA Modulation of Peritoneal Cavity Tissue-Resident Macrophages In Vivo in Mice
Mark Gurney 1, Luke C. Davies 1,2, Ruth E. Jones 1, Valentina M. Bart 1, Robert H. Jenkins 1, Paul Brennan 1, Philip R. Taylor 1,3, Magdalena A. Czubala 1
1Systems Immunity Research Institute, Division of Infection and Immunity, Cardiff University, 2Biomedical Sciences Unit, Faculty of Medicine, Health and Life Science, Swansea University, 3UK Dementia Research Institute

We demonstrate a step-by-step protocol for the investigation of gene function in peritoneal tissue-resident macrophages in vivo, using lentiviral vectors.

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Biology

Near-Infrared Spectroscopy During Reactive Hyperemia for the Assessment of Lower Limb Vascular Function
Yuri Kriel 1, Andrzej Kwintowski 1, Krist Feka 1, Mark Windsor 1, Grace Young 2, Meegan Walker 1, Christopher D. Askew 1,3
1VasoActive Research Group, School of Health, University of the Sunshine Coast, 2Centre for Trials Research, College of Biomedical and Life Sciences, Cardiff University, 3Sunshine Coast Health Institute, Sunshine Coast Hospital and Health Service

Here, we describe a non-invasive approach using near-infrared spectroscopy to assess reactive hyperemia in the lower limb. This protocol provides a standardized assessment of vascular and microvascular responsiveness that may be used to determine the presence of vascular dysfunction as well as the efficacy of therapeutic interventions.

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