Analysis of reproducibility and robustness of a renal proximal tubule microphysiological system OrganoPlate 3-lane 40 for in vitro studies of drug transport and toxicity

Abstract

Microphysiological systems are an emerging area of in vitro drug development, and their independent evaluation is important for wide adoption and use. The primary goal of this study was to test reproducibility and robustness of a renal proximal tubule microphysiological system, OrganoPlate® 3-lane 40, as an in vitro model for drug transport and toxicity studies. This microfluidic model was compared to static multi-well cultures and tested using several human renal proximal tubule epithelial cell (RPTEC) types. The model was characterized in terms of the functional transport for various tubule-specific proteins, epithelial permeability of small molecules (cisplatin, tenofovir and perfluorooctanoic acid) versus to large-molecules (fluorescent dextrans, 60-150 kDa), and gene expression response to a nephrotoxic xenobiotic. The advantages offered by OrganoPlate® 3-lane 40 as compared to multi-well cultures are presence of media flow, albeit intermittent, and increased throughput compared to other microfluidic models. However, OrganoPlate® 3-lane 40 model appeared to offer only limited (e.g., MRP-mediated transport) advantages in terms of either gene expression or functional transport when compared to the multi-well plate culture conditions. While OrganoPlate® 3-lane 40 can be used to study cellular uptake and direct toxic effects of small molecules, it may have limited utility for studies for drug transport studies. Overall, this study offers refined experimental protocols and comprehensive comparative data on the function of RPETCs in traditional multi-well culture and micro-fluidic OrganoPlate® 3-lane 40, information that will be invaluable for the prospective end-users of in vitro models of the human proximal tubule.

Characterization of Organic Anion and Cation Transport in Three Human Renal Proximal Tubular Epithelial Models

“…For instance, primary PTEC have been demonstrated to exhibit probenecid-inhibitable PAH transepithelial transport in an MPS device but not when maintained under static conditions [52]. However, other studies have shown that MPS systems did not improve OAT functionality [42,53]. Nevertheless, MPS systems could offer a promising avenue for further exploration for such studies, including improving the differentiated phenotype of iPSC-derived PTL.…”

Neuropathogenesis-on-chips for neurodegenerative diseases

“…Tissue chip testing centres established by the NIH National Center for Advancing Translational Sciences initiated grantees to validate various types of tissue chip platforms for drug testing 566 , 567 . Under these grants, three independent organisations validated microphysiological systems that replicated the kidney, liver, and BBB for drug transport and toxicity studies, with the goal of increasing the reliability, reproducibility, robustness, and throughput of its effective application 568 .…”