Sorbonne Universite 6 articles published in JoVE Developmental Biology Efficient Vascularization of Kidney Organoids through Intracelomic Transplantation in Chicken Embryos Marije Koning1,2, Ellen Lievers1,2, Thierry Jaffredo3, Cathelijne W. van den Berg1,2,4, Ton J. Rabelink1,2,4 1Department of Internal Medicine - Nephrology, Leiden University Medical Center, 2Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center, 3IBPS, CNRS UMR7622, Developmental Biology Laboratory, Sorbonne Université, 4The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center Here, we present a detailed protocol for the transplantation of kidney organoids in the celomic cavity of chicken embryos. This method induces vascularization and enhanced maturation of the organoids within 8 days and can be used to study these processes in an efficient manner. Bioengineering Reconstitution of Actin-Based Motility with Commercially Available Proteins Cécile Sykes1, Julie Plastino1 1Laboratoire de physique de l’Ecole Normale Supérieure, ENS, Université PSL, CNRS, Sorbonne Université, Université Paris Cité This protocol describes how to produce actin comets on the surfaces of beads using commercially available protein ingredients. Such systems mimic the protrusive structures found in cells, and can be used to examine physiological mechanisms of force production in a simplified way. Biology Bottom-Up In Vitro Methods to Assay the Ultrastructural Organization, Membrane Reshaping, and Curvature Sensitivity Behavior of Septins Brieuc Chauvin*1, Koyomi Nakazawa*1, Alexandre Beber1,7, Aurélie Di Cicco1, Bassam Hajj1, François Iv2, Manos Mavrakis2, Gijsje H. Koenderink3, João T. Cabral4, Michaël Trichet5, Stéphanie Mangenot*6, Aurélie Bertin*1 1Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University, Sorbonne Université, 2Institut Fresnel, CNRS UMR7249, Aix Marseille Univ, Centrale Marseille, 3Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, 4Department of Chemical Engineering, Imperial College London, 5Sorbonne Université, CNRS, Institut de Biologie Paris-Seine (IBPS), Service de microscopie électronique (IBPS-SME), 6Laboratoire Matière et Systèmes Complexes (MSC), Université Paris Cité, 7Institute of Biotechnology, Czech Academy of Sciences, BIOCEV Septins are cytoskeletal proteins. They interact with lipid membranes and can sense but also generate membrane curvature at the micron scale. We describe in this protocol bottom-up in vitro methodologies for analyzing membrane deformations, curvature-sensitive septin binding, and septin filament ultrastructure. Biochemistry Purification and Quality Control of Recombinant Septin Complexes for Cell-Free Reconstitution Gerard Castro-Linares1, Jeffrey den Haan1, Francois Iv2, Carla Silva Martins2, Aurélie Bertin3, Manos Mavrakis2, Gijsje H. Koenderink1 1Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, 2Institut Fresnel, CNRS UMR7249, Aix Marseille Univ, Centrale Marseille, 3Institut Curie, Université PSL, Sorbonne Université In vitro reconstitution of cytoskeletal proteins is a vital tool to understand the basic functional properties of these proteins. The present paper describes a protocol to purify and assess the quality of recombinant septin complexes, which play a central role in cell division and migration. Chemistry Thermochemical Studies of Ni(II) and Zn(II) Ternary Complexes Using Ion Mobility-Mass Spectrometry Anna J. Corrales1, Anna V. Arredondo1, Amber A. Flores1, Chloe L. Duvak1, Charles L. Mitchell1, Riccardo Spezia2, Laurence A. Angel1 1Department of Chemistry, Texas A&M University-Commerce, 2Laboratoire de Chimie Théorique, Sorbonne Université This article describes an experimental protocol using electrospray-ion mobility-mass spectrometry, semi-empirical quantum calculations, and energy-resolved threshold collision-induced dissociation to measure the relative thermochemistry of the dissociation of related ternary metal complexes. Biology Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe Aline Stedman1,3, Antonin Levy1,4, Philippe J. Sansonetti1,2,5, Giulia Nigro1,6 1Molecular Microbial Pathogenesis Unit, Institut Pasteur, 2Chaire de Microbiologie et Maladies Infectieuses, Collège de France, 3Institut de Biologie Paris Seine (IBPS) - Developmental Biology Unit, Sorbonne Université, CNRS UMR7622, INSERM U1156, 4Molecular Radiotherapy, INSERM U1030, Gustave Roussy, Université Paris-Saclay, 5The Center for Microbes, Development and Health, Institut Pasteur Shanghai and Chinese Academy of Sciences, 6Microenvironment and Immunity Unit, Institut Pasteur The present protocol describes a method to detect reactive oxygen species (ROS) in the intestinal murine organoids using qualitative imaging and quantitative cytometry assays. This work can be potentially extended to other fluorescent probes to test the effect of selected compounds on ROS.
