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Université de Paris

6 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Monitoring Influenza Virus Survival Outside the Host Using Real-Time Cell Analysis
Thomas Labadie 1, Quentin Grassin 2, Christophe Batéjat 2, Jean-Claude Manuguerra 2, India Leclercq 2,3
1Department of Infection Biology, London School of Hygiene and Tropical Medicine, 2Institut Pasteur, Unité Environnement et Risques Infectieux, Cellule d'Intervention Biologique d'Urgence (CIBU), 3Cellule Pasteur, Université de Paris

Reported here is a protocol for the quantification of infectious viral particles using real-time monitoring of electrical impedance of infected cells. A practical application of this method is presented by quantifying influenza A virus decay under different physicochemical parameters mimicking environmental conditions.

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Neuroscience

Whole-Brain 3D Activation and Functional Connectivity Mapping in Mice using Transcranial Functional Ultrasound Imaging
Adrien Bertolo *1, Mohamed Nouhoum *1, Silvia Cazzanelli *1, Jeremy Ferrier *3, Jean-Charles Mariani 2, Andrea Kliewer 4,2, Benoit Belliard 1, Bruno-Félix Osmanski 3, Thomas Deffieux *1, Sophie Pezet *1, Zsolt Lenkei *2, Mickael Tanter *1
1Physics for Medicine Paris, ESPCI Paris, INSERM, CNRS, PSL Research University, 2Institute of Psychiatry and Neurosciences of Paris, INSERM U1266, Université de Paris, 3Iconeus, 4Department of Pharmacology and Toxicology, Jena University Hospital - Friedrich Schiller University Jena

This protocol describes the quantification of volumetric cerebral hemodynamic variations in the mouse brain using functional ultrasound (fUS). Procedures for 3D functional activation map following sensory stimulation as well as resting-state functional connectivity are provided as illustrative examples, in anesthetized and awake mice.

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Immunology and Infection

Partial Heterotopic Hindlimb Transplantation Model in Rats
Marion Goutard 1,2,3, Mark A. Randolph 1,2,3, Corentin B. Taveau 1,2,3,4, Elise Lupon 1,2,3, Laurent Lantieri 4, Korkut Uygun 1,2,3, Curtis L. Cetrulo Jr. *1,2,3, Alexandre G. Lellouch *1,2,3,4
1Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital, Harvard Medical School, 2Vascularized Composite Allotransplantation Laboratory, Center for Transplantation Sciences, Massachusetts General Hospital, Harvard Medical School, 3Shriners Hospital for Children, 4Service de Chirurgie Plastique, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris (APHP), Université de Paris

This paper presents a partial heterotopic osteomyocutaneous flap transplantation protocol in rats and its potential outcomes in the mid-term follow-up.

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Developmental Biology

2D and 3D Human Induced Pluripotent Stem Cell-Based Models to Dissect Primary Cilium Involvement during Neocortical Development
Lucile Boutaud *1, Marie Michael *1, Céline Banal 2, Damelys Calderon 1, Sarah Farcy 1, Julie Pernelle 1, Nicolas Goudin 3, Camille Maillard 1, Clémantine Dimartino 1, Cécile Deleschaux 1, Sébastien Dupichaud 4, Corinne Lebreton 1, Sophie Saunier 1, Tania Attié-Bitach 1,5, Nadia Bahi-Buisson 1,6, Nathalie Lefort 2, Sophie Thomas 1
1Imagine Institute, INSERM UMR 1163, Université de Paris, 2Imagine Institute, iPSC Core Facility, INSERM UMR U1163, Université de Paris, 3Necker Bio-image Analysis platform of the SFR Necker, 4Imagine Institute, Cell Imaging Platform, INSERM-US24-CNRS UMS 3633 Structure Fédérative de Recherche Necker, INSERM UMR U1163, Université de Paris, 5Fédération de Génétique, Hôpital Necker-Enfants Malades, Assistance Publique Hôpitaux de Paris, 6Pediatric Neurology, APHP- Necker Enfants Malades Hospital

We present detailed protocols for the generation and characterization of 2D and 3D human induced pluripotent stem cell (hIPSC)-based models of neocortical development as well as complementary methodologies enabling qualitative and quantitative analysis of primary cilium (PC) biogenesis and function.

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Neuroscience

Magnetic Isolation of Microglial Cells from Neonate Mouse for Primary Cell Cultures
Cindy Bokobza *1, Alice Jacquens *1,2, Manuela Zinni 1, Valérie Faivre 1, Jennifer Hua 1, David Guenoun 1, Caroline Userovici 1, Shyamala Mani 1, Vincent Degos 1,2, Pierre Gressens 1, Juliette Van Steenwinckel 1
1NeuroDiderot, Inserm UMR-1141, Hôpital Robert Debré 48, Université de Paris, 2Department of anesthesia and critical care, APHP-Sorbonne university

Primary microglia cultures are commonly used to evaluate new anti-inflammatory molecules. The present protocol describes a reproducible and relevant method to magnetically isolate microglia from neonate pups.

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Biology

Primary Human Nasal Epithelial Cells: Biobanking in the Context of Precision Medicine
Mairead Kelly 1,2, Elise Dreano 1,2, Aurelie Hatton 1,2, Agathe Lepissier 1,2, Anita Golec 1,2, Isabelle Sermet-Gaudelus 1,2,3, Iwona Pranke 1,2,3
1Institut Necker Enfants Malades, 2Université de Paris, 3Centre de Référence Maladies Rares Mucoviscidose et Maladies apparentées, Assistance Publique Hôpitaux de Paris

Here we describe the isolation, amplification, and differentiation of primary human nasal epithelial (HNE) cells at the air-liquid interface and a biobanking protocol allowing to successfully freeze and then thaw amplified HNE. The protocol analyzes electrophysiological properties of differentiated HNE cells and CFTR-related chloride secretion correction upon different modulator treatments.

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