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A perfusion chamber for monitoring transepithelial NaCl transport in an in vitro model of the renal tubule
Author(s) -
Yeste Jose,
MartínezGimeno Laura,
Illa Xavi,
Laborda Pablo,
Guimerà Anton,
SánchezMarín Juan P.,
Villa Rosa,
Giménez Ignacio
Publication year - 2018
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26574
Subject(s) - reabsorption , transepithelial potential difference , tubule , biophysics , chemistry , renal physiology , ussing chamber , perfusion , kidney , proximal tubule , electrolyte , ion transporter , apical membrane , in vitro , biochemistry , medicine , endocrinology , membrane , biology , renal function , electrode
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., H 2 O 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.