Assistance Publique Hôpitaux de Paris

3 ARTICLES PUBLISHED IN JoVE

Medicine

Standardized Measurement of Nasal Membrane Transepithelial Potential Difference (NPD)

George M. Solomon¹, Inez Bronsveld², Kathryn Hayes³, Michael Wilschanski⁴, Paola Melotti⁵, Steven M. Rowe¹, Isabelle Sermet-Gaudelus^6,7

¹Department of Medicine and the Gregory Fleming James Cystic Fibrosis Center, University of Alabama at Birmingham, ²Department of Pulmonology and Tuberculosis, University Medical Center Utrecht, ³Center for Experimental Medicine, Queens University, Northern Ireland, ⁴Hadassah Hebrew University Medical Center, Jerusalem, ⁵Centro Fibrosi Cistica, Azienda Ospedaliera Universitaria Integrata, ⁶Service de Pneumologie et Allergologie Pédiatriques and Center de Ressources et de Compétence de la Mucoviscidose, Hôpital Necker Enfants Malades, ⁷INSERM U 1151, Institut Necker Enfants Malades

Here, we present a standardized protocol to measure the nasal potential difference (NPD). Cystic fibrosis transmembrane conductance regulator (CFTR) and epithelial sodium channel (ENaC) function are evaluated by the change in the voltage across the nasal epithelium after superfusion of solutions that modify ion channel activity, providing an outcome measure.

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 Banal², Damelys Calderon¹, Sarah Farcy¹, Julie Pernelle¹, Nicolas Goudin³, Camille Maillard¹, Clémantine Dimartino¹, Cécile Deleschaux¹, Sébastien Dupichaud⁴, Corinne Lebreton¹, Sophie Saunier¹, Tania Attié-Bitach^1,5, Nadia Bahi-Buisson^1,6, Nathalie Lefort², Sophie Thomas¹

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.

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.