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Ghent University Hospital

9 ARTICLES PUBLISHED IN JoVE

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Medicine

The In ovo CAM-assay as a Xenograft Model for Sarcoma
Gwen M.L. Sys 1, Lore Lapeire 2, Nikita Stevens 1, Herman Favoreel 3, Ramses Forsyth 4, Marc Bracke 2, Olivier De Wever 2
1Department of Orthopaedic Surgery and Traumatology, Ghent University Hospital, 2Department of Radiation Oncology and Experimental Cancer Research, Ghent University, 3Department of Virology, Parasitology, and Immunology, Ghent University, 4Pathlicon

The in ovo chorioallantoic membrane (CAM) is grafted with fresh sarcoma-derived tumor tissues, their single cell suspensions, and permanent and transient fluorescently labeled established sarcoma cell lines. The model is used to study graft- (viability, Ki67 proliferation index, necrosis, infiltration) and host (fibroblast infiltration, vascular ingrowth) behavior.

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Immunology and Infection

An Optimized Method for Isolating and Expanding Invariant Natural Killer T Cells from Mouse Spleen
Srinath Govindarajan 1,2, Dirk Elewaut 1,2, Michael Drennan 1,2
1Department of Rheumatology, Laboratory for Molecular Immunology and Inflammation, Ghent University Hospital, 2VIB Inflammation Research Center, Ghent University

Here we present an adapted protocol that can be used to generate a large number of murine invariant natural killer T cells from mouse spleen. The protocol outlines an approach by which splenic iNKT cells can be enriched for, isolated and expanded in vitro using a limited number of animals and reagents.

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JoVE Journal

Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange
Tim Pieters 1,2,3,4, Lieven Haenebalcke 1,2, Kenneth Bruneel 1,2,4, Niels Vandamme 1,2,4, Tino Hochepied 1,2, Jolanda van Hengel 5, Dagmar Wirth 6, Geert Berx 1,2,4, Jody J. Haigh 7, Frans van Roy 1,2,4, Steven Goossens 1,2,3,4
1Department of Biomedical Molecular Biology, Ghent University, 2Inflammation Research Center, VIB, 3Center for Medical Genetics, Ghent University Hospital, 4Cancer Research Institute Ghent (CRIG), 5Department of Basic Medical Sciences, Faculty of Medicine and Health Sciences, Ghent University, 6Helmholtz Center for Infection Research, 7Mammalian Functional Genetics Laboratory, Division of Blood Cancers, Australian Centre for Blood Diseases, Department of Clinical Haematology, Monash University and Alfred Health Alfred Centre

Proteins often contain multiple domains that can exert different cellular functions. Gene knock-outs (KO) do not consider this functional diversity. Here, we report a recombination-mediated cassette exchange (RMCE)-based structure-function approach in KO embryonic stem cells that allows for the molecular dissection of various functional domains or variants of a protein.

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Cancer Research

PET and MRI Guided Irradiation of a Glioblastoma Rat Model Using a Micro-irradiator
Julie Bolcaen 1, Benedicte Descamps 2, Tom Boterberg 3, Christian Vanhove 2, Ingeborg Goethals 1
1Department of Nuclear Medicine, Ghent University Hospital, 2IBiTech-MEDISIP, Department of Electronics and Information Systems, Ghent University, 3Department of Radiation Oncology, Ghent University Hospital

In the past, small animal irradiation was usually performed without the ability to target a well-delineated tumor volume. The goal was to mimic the treatment of human glioblastoma in rats. Using a small animal irradiation platform, we performed MRI-guided 3D conformal irradiation with PET-based sub-volume boosting in a preclinical setting.

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Medicine

Vessel-sparing Excision and Primary Anastomosis
Wesley Verla 1, Willem Oosterlinck 1, Marjan Waterloos 1, Nicolaas Lumen 1
1Department of Urology, Ghent University Hospital

Here, we present an elaborate and efficient protocol to treat isolated short bulbar or posterior urethral strictures with vessel-sparing excision and primary anastomosis.

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Cancer Research

Positron Emission Tomography-based Dose Painting Radiation Therapy in a Glioblastoma Rat Model using the Small Animal Radiation Research Platform
Sam Donche 1, Jeroen Verhoeven 1, Benedicte Descamps 2, Charlotte Bouckaert 3, Robrecht Raedt 3, Christian Vanhove 2, Ingeborg Goethals 1
1Department of Diagnostic Sciences, Ghent University, 2Department of Electronics and Information Systems, Ghent University, 3Department of Head and Skin, Ghent University

Here we present a protocol to perform preclinical positron emission tomography-based radiotherapy in a rat glioblastoma model using algorithms developed in-house to optimize the accuracy and efficiency.

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Cancer Research

An Automated Microscopic Scoring Method for the γ-H2AX Foci Assay in Human Peripheral Blood Lymphocytes
Shankari Nair 1, Samantha Cairncross 2, Xanthene Miles 1, Monique Engelbrecht 3, Peter du Plessis 1, Julie Bolcaen 1, Randall Fisher 1, Roya Ndimba 1, Charnay Cunningham 4, Wilner Martínez-López 5, Christian Schunck 6, Charlot Vandevoorde 1
1Radiation Biophysics Division, Nuclear Medicine Department, iThemba LABS, 2Department of Biotechnology, University of the Western Cape, 3Department of Medical Biosciences, University of the Western Cape, 4Division of Medical Physiology, Department of Medicine and Health Sciences, Stellenbosch University, 5Biodosimetry Service, Instituto de Investigaciones Biológicas Clemente Estable and Academic Unit on Radiation Protection, Faculty of Medicine, University of the Republic, 6MetaSystems Hard & Software GmbH

This protocol presents the slide preparation and automated scoring of the γ-H2AX foci assay on peripheral blood lymphocytes. To illustrate the method and sensitivity of the assay, isolated lymphocytes were irradiated in vitro. This automated method of DNA DSB detection is useful for fast and high-throughput biological dosimetry applications.

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Medicine

Retzius-Sparing Robot-Assisted Radical Prostatectomy
Nicolaas Lumen *1, Edward Lambert *1, Filip Poelaert 1, Michel Wirtz 1, Sofie Verbeke 2, Charles Van Praet 1
1Dept. of Urology, Ghent University Hospital, 2Dept. of Pathology, Ghent University Hospital

Robot-assisted Retzius-sparing radical prostatectomy is a technique that enables to preserve urinary continence or facilitates recovery of urinary continence in the majority of patients. Patients must be informed about the risk of a positive surgical margin.

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Developmental Biology

Human Ovarian Surface Epithelium Organoids as a Platform to Study Tissue Regeneration
Julieta S. Del Valle 1,2, Azra Husetic *1, Dina Diek *1, Laurens F. Rutgers *1, Joyce D. Asseler 3,4,5, Jeroen Metzemaekers 6, Norah M. van Mello 3,4,5, Susana M. Chuva de Sousa Lopes 1,2,7
1Department of Anatomy and Embryology, Leiden University Medical Center, 2The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, 3Department of Obstetrics and Gynaecology, Amsterdam University Medical Center, 4Centre of Expertise on Gender Dysphoria, Amsterdam UMC, 5Amsterdam Reproduction and Development Research Institute, 6Department of Gynaecology, Leiden University Medical Center, 7Ghent-Fertility and Stem Cell Team (G-FAST), Department of Reproductive Medicine, Ghent University Hospital

This protocol describes establishing three-dimensional (3D) tissue organoids from primary human ovarian surface epithelium (hOSE) cells. The protocol includes isolation of hOSE from freshly collected ovaries, cellular expansion of the hOSE, cryopreservation-thawing procedures, and organoid derivation. Immunofluorescence, quantitative analysis, and showcasing utility as a screening platform are included.

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