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Hospices Civils de Lyon

7 ARTICLES PUBLISHED IN JoVE

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Medicine

Microarray-based Identification of Individual HERV Loci Expression: Application to Biomarker Discovery in Prostate Cancer
Philippe Pérot 1,2, Valérie Cheynet 1,2, Myriam Decaussin-Petrucci 1,3,4, Guy Oriol 1,2, Nathalie Mugnier 5, Claire Rodriguez-Lafrasse 1,4,6, Alain Ruffion 1,4,7, François Mallet 1,2
1Joint Unit Hospices de Lyon-bioMérieux, 2Medical Diagnostic Discovery Department, BioMérieux, 3Department of Pathology and Cytology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon, 4Medical Faculty, Lyon 1 University, 5Data and Knowledge Laboratory, BioMérieux, 6Department of Biochemistry and Molecular Biology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon, 7Department of Urology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon

Human endogenous retroviruses (HERV), which occupy 8% of the human genome, retain scarce coding capacities but a hundred thousand long terminal repeats (LTRs). A custom Affymetrix microarray was designed to identify individual HERV locus expression and was used on prostate cancer tissues as a proof of concept for future clinical studies.

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Neuroscience

Use of pHluorin to Assess the Dynamics of Axon Guidance Receptors in Cell Culture and in the Chick Embryo
Céline Delloye-Bourgeois *1, Arnaud Jacquier *1, Julien Falk 1, Valérie Castellani 1
1University Claude Bernard Lyon1, CGphiMC UMR CNRS 5534, University of Lyon

We describe here the use of a pH-sensitive green fluorescent protein variant, pHluorin, to study the spatio-temporal dynamics of axon guidance receptors trafficking at the cell surface. The pHluorin-tagged receptor is expressed both in cell culture and in vivo, using electroporation of the chick embryo.

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Medicine

Confocal Time Lapse Imaging as an Efficient Method for the Cytocompatibility Evaluation of Dental Composites
Ghania Nina Attik 1, Kerstin Gritsch 1,2, Pierre Colon 1,3, Brigitte Grosgogeat 1,2
1Laboratoire des Multimatériaux et Interfaces, UMR CNRS 5615, Université Lyon1, 2UFR d'Odontologie, Université Lyon1; Service de Consultations et de Traitements Dentaires, Hospices Civils de Lyon, 3UFR d'Odontologie, Université Paris Diderot; Service d'Odontologie, APHP, Hôpital Rothschild

The most studied aspect of the biocompatibility of dental composites is their cytotoxicity 3D CLSM time lapse imaging combined with fluorescent signal quantification allows sensitive evaluation of the cell fate over time. Utilizing this method could be an efficient tool to assess the cytocompatibility of dental composites.

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Medicine

Isolation and Characterization of a Head and Neck Squamous Cell Carcinoma Subpopulation Having Stem Cell Characteristics
Marion Gilormini 1, Anne-Sophie Wozny 1,2, Priscillia Battiston-Montagne 1, Dominique Ardail 1,2, Gersende Alphonse 1,2, Claire Rodriguez-Lafrasse 1,2
1UCBL1, UMR/CNRS 5822, Laboratoire de Radiobiologie Cellulaire Moléculaire, Université de Lyon, 2Hospices-Civils-de-Lyon, Centre Hospitalier Lyon-Sud

Understanding the role of cancer stem-like cells in tumor recurrence and resistance to therapies has become a topic of great interest in the last decade. This article describes the isolation and characterization of the sub-population of cancer stem-like cells from head and neck squamous carcinoma cell lines (HNSCC).

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Cancer Research

Evaluation of the Cell Invasion and Migration Process: A Comparison of the Video Microscope-based Scratch Wound Assay and the Boyden Chamber Assay
Jean-Baptiste Guy *1,2, Sophie Espenel *1,2, Alexis Vallard 1,2, Priscillia Battiston-Montagne 1, Anne-Sophie Wozny 1,3, Dominique Ardail 1,3, Gersende Alphonse 1,3, Chloé Rancoule 1,2, Claire Rodriguez-Lafrasse 1,3, Nicolas Magne 1,2
1UMR CNRS 5822 /IN2P3, IPNL, PRISME, Laboratoire de Radiobiologie Cellulaire et Moléculaire, Faculté de Médecine Lyon-Sud, Université Lyon 1, 2Département de Radiothérapie, Institut de Cancérologie de la Loire - Lucien Neuwirth, 3Hospices Civils de Lyon, Centre Hospitalier Lyon-Sud

This study reports two different methods for the analysis of cell invasion and migration: the Boyden chamber assay and the in vitro video microscope-based wound-healing assay. The protocols for these two experiments are described, and their benefits and disadvantages are compared.

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Neuroscience

Modeling Charcot-Marie-Tooth Disease In Vitro by Transfecting Mouse Primary Motoneurons
Arnaud Jacquier 1,2, Valérie Risson 1, Laurent Schaeffer 1,3
1Institut NeuroMyoGène, CNRS UMR5310, INSERM U1217, Université Lyon, 2Unité Fonctionnelle de Neurogénétique Moléculaire, Hospices Civils de Lyon, 3Centre de Biotechnologie Cellulaire, Hospices Civils de Lyon

The goal of this technique is to prepare a highly enriched culture of primary motoneurons (MNs) from murine spinal cord. To evaluate the consequences of mutations causing MN diseases, we describe here the isolation of these isolated MNs and their transfection by magnetofection.

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Cancer Research

Semi-automatic PD-L1 Characterization and Enumeration of Circulating Tumor Cells from Non-small Cell Lung Cancer Patients by Immunofluorescence
Jessica Garcia 1,2,3, David Barthelemy 1,2,3, Florence Geiguer 1,2, Julie Ballandier 1,2,3, Kathryn W. Li 4, Jean-Phillippe Aurel 4, Frédérique Le Breton 5, Claire Rodriguez-Lafrasse 1, Brigitte Manship 3, Sébastien Couraud 6,7, Léa Payen 1,2,3
1Laboratoire de Biochimie et Biologie Moléculaire, Groupe Hospitalier Sud, Hospices Civils de Lyon, 2Circulating Cancer (CIRCAN) Program, Hospices Civils de Lyon Cancer Institute, 3University of Lyon, Claude Bernard University, Cancer Research Center of Lyon, 4Biolidics Limited, 5Institut de Pathologie Multisites des HCL-Site Sud, Hospices Civils de Lyon, 6Acute Respiratory Disease and Thoracic Oncology Department, Lyon Sud Hospital, Hospices Civils de Lyon Cancer Institute, 7EMR-3738 Therapeutic Targeting in Oncology, Lyon Sud Medical Faculty, Lyon 1 University

The characterization of circulating tumor cells (CTCs) is a popular topic in translational research. This protocol describes a semi-automatic immunofluorescence (IF) assay for PD-L1 characterization and enumeration of CTCs in non-small cell lung cancer (NSCLC) patient samples.

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