Titre
A primary culture system of mouse thick ascending limb cells with preserved function and uromodulin processing.
Type
article
Institution
UNIL/CHUV/Unisanté + institutions partenaires
Auteur(s)
Glaudemans, B.
Auteure/Auteur
Terryn, S.
Auteure/Auteur
Gölz, N.
Auteure/Auteur
Brunati, M.
Auteure/Auteur
Cattaneo, A.
Auteure/Auteur
Bachi, A.
Auteure/Auteur
Al-Qusairi, L.
Auteure/Auteur
Ziegler, U.
Auteure/Auteur
Staub, O.
Auteure/Auteur
Rampoldi, L.
Auteure/Auteur
Devuyst, O.
Auteure/Auteur
Liens vers les personnes
Liens vers les unités
ISSN
1432-2013
Statut éditorial
Publié
Date de publication
2014
Volume
466
Numéro
2
Première page
343
Dernière page/numéro d’article
356
Peer-reviewed
Oui
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov'tPublication Status: ppublish
Résumé
The epithelial cells lining the thick ascending limb (TAL) of the loop of Henle perform essential transport processes and secrete uromodulin, the most abundant protein in normal urine. The lack of differentiated cell culture systems has hampered studies of TAL functions. Here, we report a method to generate differentiated primary cultures of TAL cells, developed from microdissected tubules obtained in mouse kidneys. The TAL tubules cultured on permeable filters formed polarized confluent monolayers in ∼12 days. The TAL cells remain differentiated and express functional markers such as uromodulin, NKCC2, and ROMK at the apical membrane. Electrophysiological measurements on primary TAL monolayers showed a lumen-positive transepithelial potential (+9.4 ± 0.8 mV/cm(2)) and transepithelial resistance similar to that recorded in vivo. The transepithelial potential is abolished by apical bumetanide and in primary cultures obtained from ROMK knockout mice. The processing, maturation and apical secretion of uromodulin by primary TAL cells is identical to that observed in vivo. The primary TAL cells respond appropriately to hypoxia, hypertonicity, and stimulation by desmopressin, and they can be transfected. The establishment of this primary culture system will allow the investigation of TAL cells obtained from genetically modified mouse models, providing a critical tool for understanding the role of that segment in health and disease.
PID Serval
serval:BIB_D13D89DDAFF4
PMID
Date de création
2016-10-10T09:25:57.095Z
Date de création dans IRIS
2025-05-20T23:55:54Z