Understanding the active transport of substrates by the kidney in the renal proximal convoluted tubule is crucial for drug
development and for studying kidney diseases. Currently, cell-based assays are applied for this this purpose, however, diferences between assays and the body are common, indicating the importance of in vitro–in vivo discrepancies. Several studies
have suggested that 3D cell cultures expose cells to a more physiological environments, thus, providing more accurate cell
function results. To mimic the renal proximal tubule, we have developed a custom-made renal module (RM), containing a
single polypropylene hollow fbre (Plasmaphan P1LX, 3M) that serves as a porous scafold and compared to conventional
Transwell cell-based bidirectional transport studies. In addition, a constant fow of media, exposed cells to a physiological
shear stress of 0.2 dyne/cm2
. MDCK-Mdr1a cells, overexpressing the rat Mdr1a (P-gp) transporter, were seeded onto the
HF membrane surface coated with the basement membrane matrix Geltrex which facilitated cell adhesion and tight junction formation. Cells were then seeded into the HF lumen where attachment and tight junction formation were evaluated
by fuorescence microscopy while epithelial barrier integrity under shear stress was shown to be achieved by day 7. qPCR
results have shown signifcant changes in gene expression compared to cells grown on Transwells. Kidney injury marker
such as KIM-1 and the hypoxia marker CA9 have been downregulated, while the CD133 (Prominin-1) microvilli marker has
shown a fvefold upregulation. Furthermore, the renal transporter P-gp expression has been downregulated by 50%. Finally,
bidirectional assays have shown that cells grown in the RM were able to reabsorb albumin with a higher efciency compared
to Transwell cell cultures while efux of the P-gp-specifc substrates Hoechst and Rhodamine 123 was decreased. These
results further support the efect of the microenvironment and fuidic shear stress on cell function and gene expression. This
can serve as the basis for the development of a microphysiological renal model for drug transport studies.
Funding
Creation of bioartificial kidney with renal cells primary immortalised or stem as a model of renal transport
Biotechnology and Biological Sciences Research Council
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