AP-HP

6 articles published in JoVE

• Biology

  
  Unfractionated Bulk Culture of Mouse Skeletal Muscle to Recapitulate Niche and Stem Cell Quiescence Louai Zaidan*¹, Perla Geara*¹, Matthew J. Borok*¹, Léo Machado¹, Despoina Mademtzoglou¹, Philippos Mourikis¹, Frederic Relaix^{1,2,3,4 1}Univ Paris Est Creteil, INSERM, IMRB, F-94010 Creteil, France, ²Ecole nationale vétérinaire d'Alfort, IMRB, F-94700 Maisons-Alfort, France, ³EFS, IMRB, F-94010 Creteil, France, ⁴AP-HP, Hopital Mondor, Service d'histologie, F-94010 Creteil, France Skeletal muscle comprises multiple cell types, including resident stem cells, each with a special contribution to muscle homeostasis and regeneration. Here, the 2D culture of muscle stem cells and the muscle cell niche in an ex vivo setting that preserves many of the physiological, in vivo, and environmental characteristics are described.

• Immunology and Infection

  
  Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses Camille Bouillier¹, Vincent Rincheval¹, Delphine Sitterlin¹, Sabine Blouquit-Laye¹, Aurore Desquesnes¹, Jean-François Eléouët², Elyanne Gault^1,3, Marie-Anne Rameix-Welti^{1,3 1}UMR 1173 Institut national de la santé et de la recherche médicale (INSERM), Université de Versailles St. Quentin, ²UR892 Institut national de la recherche agronomique (INRA), Unité de virologie et immunologie moléculaires, ³Assistance Publique–Hôpitaux de Paris (AP-HP), Hôpital Ambroise Paré, Laboratoire de Microbiologie We describe a method for generating and amplifying genetically modified respiratory syncytial viruses (RSVs) and an optimized plaque assay for RSVs. We illustrate this protocol by creating two recombinant viruses that respectively allow quantification of RSV replication and live analysis of RSV inclusion bodies and inclusion bodies-associated granules dynamics.

• Cancer Research

  
  Immunoglobulin Gene Sequence Analysis In Chronic Lymphocytic Leukemia: From Patient Material To Sequence Interpretation Andreas Agathangelidis*¹, Lesley Ann Sutton*^2,3, Anastasia Hadzidimitriou¹, Cristina Tresoldi⁴, Anton W. Langerak⁵, Chrysoula Belessi⁶, Frederic Davi⁷, Richard Rosenquist^2,3, Kostas Stamatopoulos^1,2, Paolo Ghia^{8 1}Institute of Applied Biosciences, Centre for Research and Technology Hellas, ²Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, ³Department of Molecular Medicine and Surgery, Karolinska Institutet, ⁴Division of Immunology, Transplantation and Infectious, IRCCS San Raffaele Scientific Institute, ⁵Department of Immunology, Laboratory for Medical Immunology, Erasmus University Medical Center, ⁶Hematology Department, Nikea General Hospital, ⁷Assistance publique - Hôpitaux de Paris (AP-HP), Hopital Pitié-Salpêtrière, Department of Hematology, and UPMC University Paris 06, UMRS 1138, ⁸Division of Experimental Oncology, IRCCS Istituto Scientifico San Raffaele and Università Vita-Salute San Raffaele Herein, we present a protocol that details the technical aspects and essential requirements to ensure robust IG gene sequence analysis in patients with chronic lymphocytic leukemia (CLL), based on the accumulated experience of the European Research initiative on CLL (ERIC).

• Developmental Biology

  
  Engineering Transplantation-suitable Retinal Pigment Epithelium Tissue Derived from Human Embryonic Stem Cells Karim Ben M'Barek^1,2,3, Walter Habeler^1,2,3, Alexandra Plancheron^1,2,3, Mohamed Jarraya⁴, Olivier Goureau⁵, Christelle Monville^{1,2 1}U861, I-Stem, Association Française contre les Myopathies (AFM), Institut National de la Santé et de la Recherche Médicale (INSERM), ²I-Stem, Association Française contre les Myopathies (AFM), Centre pour L’Etude des Cellules Souches (CECS), ⁴Banque de tissus humain, Hôpital Saint Louis, Assistance Publique - Hôpitaux de Paris (AP-HP), ⁵Sorbonne Université, INSERM, CNRS, Institut de la Vision, F-75012 We describe a method to engineer a retinal tissue composed of retinal pigment epithelial cells derived from human pluripotent stem cells cultured on top of human amniotic membranes and its preparation for grafting in animal models.

• Immunology and Infection

  
  Analysis of Microglia and Monocyte-derived Macrophages from the Central Nervous System by Flow Cytometry Elodie Martin^1,2,3, Mohamed El-Behi^1,2,3, Bertrand Fontaine^1,2,3,4, Cecile Delarasse^{1,2,3 1}Inserm U 1227, CNRS UMR 7225, ²Sorbonne Universités, UPMC, University of Paris, ³Institut du Cerveau et de la Moelle épinière, ICM, ⁴AP-HP, Hôpital de la Pitié Salpêtrière; This protocol provides an analysis of the macrophage subpopulations in the adult mouse central nervous system by flow cytometry and is helpful for the study of multiple markers expressed by these cells.

• Medicine

  
  Continuous Manual Exchange Transfusion for Patients with Sickle Cell Disease: An Efficient Method to Avoid Iron Overload Bérengère Koehl¹, Florence Missud¹, Laurent Holvoet¹, Ghislaine Ithier¹, Oliver Sakalian-Black², Zinedine Haouari¹, Emmanuelle Lesprit³, André Baruchel⁴, Malika Benkerrou^{1 1}Reference Centre of Sickle Cell Disease, Hematology Unit, Robert Debré Hospital, AP-HP, ²School of Pharmacy, Université Paris Descartes, ³Etablissement Français du Sang, Robert Debré Hospital, AP-HP, ⁴Hematology Unit,, Robert Debré Hospital, AP-HP, Univsité Paris Diderot We have outlined a method of continuous manual exchange transfusion for the treatment of sickle cell disease in patients. This safe protocol was designed to effectively limit iron overload in patients in need of chronic transfusions and can be used extensively without any special equipment.