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Université Lyon 1

6 ARTICLES PUBLISHED IN JoVE

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Biology

Isolation and Culture of Mouse Primary Pancreatic Acinar Cells
Johann Gout 1,2,3,4,5, Roxane M. Pommier 1,2,3,4,5, David F. Vincent 1,2,3,4,5, Bastien Kaniewski 1,2,3,4,5, Sylvie Martel 1,2,3,4,5, Ulrich Valcourt *1,2,3,4,5, Laurent Bartholin *1,2,3,4,5
1INSERM U1052, Centre de Recherche en Cancérologie de Lyon, 2CNRS UMR5286, Centre de Recherche en Cancérologie de Lyon, 3Université de Lyon, 4Université Lyon 1, 5Centre Léon Bérard

In this publication, we describe a rapid and convenient procedure for isolating and culturing primary pancreatic acinar cells from the murine pancreas. This method constitutes a valuable approach to study the physiology of fresh primary normal/untransformed exocrine pancreatic cells.

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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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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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Chemistry

Analysis of Minerals Produced by hFOB 1.19 and Saos-2 Cells Using Transmission Electron Microscopy with Energy Dispersive X-ray Microanalysis
Lukasz Bozycki 1, Magdalena Komiazyk 1, Saida Mebarek 2,3,4,5,6, Rene Buchet 2,3,4,5,6, Slawomir Pikula 1, Agnieszka Strzelecka-Kiliszek 1
1Laboratory of Lipid Biochemistry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, 2Université de Lyon, 3Université Lyon 1, 4L'insitut National des Sciences Appliquées (INSA) de Lyon, 5Ecole Supérieure de Chimie Physique Electronique (CPE) Lyon, 6Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche (UMR), L'institut de Chimie et Biochimie Moléculaires et Supramoléculaires (ICBMS)

We present a protocol to compare the state of minerals in vesicles released by two human bone cell lines: hFOB 1.19 and Saos-2. Their mineralization profiles were analyzed by Alizarin Red-S (AR-S) staining, ultraviolet (UV) light visualization, transmission electron microscopy (TEM) imaging and energy dispersive X-ray microanalysis (EDX).

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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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