Université Lyon 1

6 ARTICLES PUBLISHED IN JoVE

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.

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 Mugnier⁵, 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.

Medicine

Isolation and Characterization of a Head and Neck Squamous Cell Carcinoma Subpopulation Having Stem Cell Characteristics

Marion Gilormini¹, Anne-Sophie Wozny^1,2, Priscillia Battiston-Montagne¹, 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).

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-Montagne¹, 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.

Chemistry

Analysis of Minerals Produced by hFOB 1.19 and Saos-2 Cells Using Transmission Electron Microscopy with Energy Dispersive X-ray Microanalysis

Lukasz Bozycki¹, Magdalena Komiazyk¹, Saida Mebarek^2,3,4,5,6, Rene Buchet^2,3,4,5,6, Slawomir Pikula¹, Agnieszka Strzelecka-Kiliszek¹

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).

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. Li⁴, Jean-Phillippe Aurel⁴, Frédérique Le Breton⁵, Claire Rodriguez-Lafrasse¹, Brigitte Manship³, 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.