Universite de Lyon

11 articles published in JoVE

• Biology

  
  A Human Bone Marrow 3D Model to Investigate the Dynamics and Interactions Between Resident Cells in Physiological or Tumoral Contexts Kawtar Arizkane^1,2,3, Kevin Geistlich^1,2,3,4, Laurine Moindrot^1,2,3, Emma Risson^1,2,3, Sandrine Jeanpierre^1,2,3,4, Léa Barral^1,2,3, Anaëlle Bobard^1,2,3, Giulio Menegazzi⁵, Thibault Voeltzel^1,2,3, Véronique Maguer-Satta^1,2,3, Sylvain Lefort^{1,2,3 1}Centre de Recherche en Cancérologie de Lyon, ²Centre de Recherche en Cancérologie de Lyon, ³Université de Lyon, ⁴Centre Léon Bérard, ⁵Department of Experimental and Clinical Biomedical Sciences Mario Serio, Università degli Studi di Firenze Here, we describe an easy-to-implement, standardized, microphysiological system that reflects the complexity of the human bone marrow's in vivo structure, providing a pertinent model to finely study a broad range of normal and pathological events.

• Biology

  
  Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds Josselin Rigaill*^1,2, Estelle Audoux*¹, Killian Rodriguez¹, Aurélien Peyron¹, Philippe Berthelot^1,3, Jérôme Josse^4,5, Frédéric Laurent^4,5, Robin Caire¹, Paul O. Verhoeven^{1,2 1}CIRI, Centre International de Recherche en Infectiologie, GIMAP team, University of Lyon, INSERM U1111, CNRS, UMR5308, ENS Lyon, UCBL1, University of St-Etienne, France, ²Department of Infectious Agents and Hygiene, University Hospital of St-Etienne, St-Etienne, France, ³Department of Infectious Diseases, University Hospital of St-Etienne, St-Etienne, France, ⁴CIRI, Centre International de Recherche en Infectiologie, Staphylococcal Pathogenesis team, University of Lyon, INSERM U1111, CNRS UMR5308, ENS Lyon, UCBL1, University of Lyon, Lyon, France, ⁵Department of Bacteriology, Institute for Infectious Agents, Hospices Civils de Lyon, Lyon, France This protocol aims to describe how to study the extent of Staphylococcus aureus internalization and its ability to survive inside the human host cell, as well as the intracellular efficacy of antimicrobial compounds.

• Cancer Research

  
  A Three-dimensional Model of Spheroids to Study Colon Cancer Stem Cells Maria Virginia Giolito^1,2, Léo Claret^1,2, Carla Frau¹, Michelina Plateroti^{1,2 1}Département de la recherche, Centre de Recherche en Cancérologie de Lyon, INSERM U1052, CNRS UMR5286, Université de Lyon, Université Lyon 1, Centre Léon Bérard, ²UMR-S1113 - IRFAC INSERM, Université de Strasbourg This protocol presents a novel, robust, and reproducible culture system to generate and grow three-dimensional spheroids from Caco2 colon adenocarcinoma cells. The results provide the first proof-of-concept for the appropriateness of this approach to study cancer stem cell biology, including the response to chemotherapy.

• Developmental Biology

  
  Use of Atomic Force Microscopy to Measure Mechanical Properties and Turgor Pressure of Plant Cells and Plant Tissues Simone Bovio^1,2, Yuchen Long², Françoise Monéger^{2 1}SFR Biosciences, Université de Lyon, ²Laboratoire de Reproduction et Développement des Plantes, Université de Lyon, ENS de Lyon, UCBL, INRA, CNRS Here, we present atomic force microscopy (AFM), operated as a nano- and micro-indentation tool on cells and tissues. The instrument allows the simultaneous acquisition of 3D surface topography of the sample and its mechanical properties, including cell wall Young's modulus as well as turgor pressure.

• 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-Kiliszek^{1 1}Laboratory of Lipid Biochemistry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, ²Université de Lyon, ³Université Lyon 1, ⁴ 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).

• Biology

  
  Hemi-laryngeal Setup for Studying Vocal Fold Vibration in Three Dimensions Christian T. Herbst^1,2, Vit Hampala¹, Maxime Garcia^2,3, Riccardo Hofer², Jan G. Svec^{1 1}Voice Research Lab, Department of Biophysics, Faculty of Science, Palacky University Olomouc, ²Laboratory of Bio-Acoustics, Dept. of Cognitive Biology, University of Vienna, ³ENES Lab, NEURO-PSI,CNRS UMR 9197, Université Lyon/Saint-Etienne, France This paper introduces a protocol for the preparation of hemi-larynx specimens facilitating a multi-dimensional view of vocal fold vibration, in order to investigate various biophysical aspects of voice production in humans and non-human mammals.

• Biology

  
  Glucose Uptake Measurement and Response to Insulin Stimulation in In Vitro Cultured Human Primary Myotubes Stephanie Chanon¹, Christine Durand¹, Aurelie Vieille-Marchiset¹, Maud Robert², Charna Dibner³, Chantal Simon¹, Etienne Lefai^{1 1}CarMeN Laboratory, INSERM U1060, INRA 1397, University of Lyon, ²Department of digestive and bariatric surgery, Obesity Integrated Center, University Hospital of Edouard Herriot, Hospices Civils de Lyon, Lyon 1 University, ³Division of Endocrinology, Diabetes, Hypertension and Nutrition, Department of Clinical Medicine, Faculty of Medicine, University of Geneva In this method, human primary muscle cells are cultured in vitro to obtain differentiated myotubes and glucose uptake rates are measured. We provide a detailed protocol to quantify rates in basal and insulin-stimulated states using radiolabeled [³H] 2-deoxy-D-Glucose.

• 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 1}UCBL1, 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).

• 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 1}INSERM 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.

• Bioengineering

  
  Engineering Adherent Bacteria by Creating a Single Synthetic Curli Operon Benoît Drogue¹, Philippe Thomas², Laurent Balvay³, Claire Prigent-Combaret¹, Corinne Dorel^{4 1}UMR CNRS 5557 Ecologie Microbienne, Université Lyon 1, Université de Lyon, ²Département Biosciences, INSA de Lyon, Université de Lyon, ³INSERM U758, Ecole Normale Supérieure de Lyon, Université de Lyon, ⁴Laboratoire de Génie Civil et Ingénierie Environnementale, INSA de Lyon, Université de Lyon The design of a synthetic operon encoding both the secretory apparatus and the structural monomers of curli fibers is described. Overproduction of these amyloids and adherent polymers allows a measurable gain of adherence of the E. coli chassis¹. Easy ways to visualize and quantify adherence are explained.

• Bioengineering

  
  Improved Visualization and Quantitative Analysis of Drug Effects Using Micropatterned Cells Sébastien Degot¹, Muriel Auzan¹, Violaine Chapuis¹, Anne Béghin², Amélie Chadeyras¹, Constantin Nelep¹, Maria Luisa Calvo-Muñoz¹, Joanne Young¹, François Chatelain¹, Alexandra Fuchs^{1 1}CYTOO Cell Architects, Grenoble, France, ²Centre Commun de Quantimétrie, Faculté de Médecine Rockefeller, Lyon, France Adhesive micropatterns that normalize cellular architecture can be used to increase sensitivity in the detection of drug effects, improve reproducibility and simplify automated image acquisition and analysis. Such technology will benefit drug/siRNA screening assays, performed on conventional cell culture supports and consequently suffering from excessive cell-to-cell variability.