A perfusion chamber for monitoring transepithelial NaCl transport in an in vitro model of the renal tubule

Yeste, J. ; Martínez-Gimeno, L. ; Illa, X. ; Laborda, P. ; Guimerà, A. ; Sánchez-Marín, J.P. ; Villa, R. ; Giménez, I. (Universidad de Zaragoza)

Resumen: Transepithelial electrical measurements in the renal tubule have provided a better understanding of how kidney regulates electrolyte and water homeostasis through the reabsorption of molecules and ions (e.g., H2O and NaCl). While experiments and measurement techniques using native tissue are difficult to prepare and to reproduce, cell cultures conducted largely with the Ussing chamber lack the effect of fluid shear stress which is a key physiological stimulus in the renal tubule. To overcome these limitations, we present a modular perfusion chamber for long-term culture of renal epithelial cells under flow that allows the continuous and simultaneous monitoring of both transepithelial electrical parameters and transepithelial NaCl transport. The latter is obtained from electrical conductivity measurements since Na+ and Cl- are the ions that contribute most to the electrical conductivity of a standard physiological solution. The system was validated with epithelial monolayers of raTAL and NRK-52E cells that were characterized electrophysiologically for 5 days under different flow conditions (i.e., apical perfusion, basal, or both). In addition, apical to basal chemical gradients of NaCl (140/70 and 70/140 mM) were imposed in order to demonstrate the feasibility of this methodology for quantifying and monitoring in real time the transepithelial reabsorption of NaCl, which is a primary function of the renal tubule.

Idioma: Inglés

DOI: 10.1002/bit.26574

Año: 2018

Publicado en: BIOTECHNOLOGY AND BIOENGINEERING 115, 6 (2018), 1604-1613
ISSN: 0006-3592

Factor impacto JCR: 4.26 (2018)

Categ. JCR: BIOTECHNOLOGY & APPLIED MICROBIOLOGY rank: 30 / 162 = 0.185 (2018) - Q1 - T1

Factor impacto SCIMAGO: 1.309 - Applied Microbiology and Biotechnology (Q1) - Biotechnology (Q1) - Bioengineering (Q1)

Financiación: info:eu-repo/grantAgreement/ES/MINECO/DPI2011-28262-C04-02
Financiación: info:eu-repo/grantAgreement/ES/MINECO/DPI2015-65401-C3-3-R
Financiación: info:eu-repo/grantAgreement/ES/MINECO/SAF2014-62114-EXP

Tipo y forma: Article (PostPrint)

Área (Departamento): Área Fisiología (Dpto. Farmacología y Fisiolog.)