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Institut Necker Enfants Malades

2 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.

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.