Background and Purpose: The kidneys are crucial for eliminating drugs and chemicals from the body, with renal epithelial cells being particularly vulnerable to damage by xenobiotics and their metabolites. Complex in vitro models of the kidney, including Microphysiological Systems (MPS), can be used to faithfully predict clinical response to toxicants. However, their high cost and low throughput often limit their application in broad contexts of use, and constant technological development complicates the process of model selection. Methods: This study compared effects of 16 compounds in concentration response design using the TERT1-OAT1 renal proximal tubule epithelial cell (RPTEC) line cultured on four platforms: standard 96- well plates, static and fluidic Transwells (PhysioMimix® T12), and OrganoPlate® 3-lane 40. Cytotoxicity was monitored via LDH leakage activity and transepithelial electrical resistance (TEER, in Transwell-based models). Samples were collected after 2, 24, and 48 hours of exposure, and drug transport was measured by LC-MS/ MS. Additionally, transcriptomic analysis was performed using the TempO-seq® S1500+ assay suite. Results: Cytotoxicity data showed that while RPTEC cultured in 96-well plates showed the lowest sensitivity, comparable effects were observed in the other platforms. When points of departure were derived comparing model predictions of human nephrotoxicity, we found that platforms incorporating fluid flow were more accurate, particularly in their sensitivity to detect “true positives”. Drug transport analysis, conducted on the barrier models (static and fluidic Transwells, and OrganoPlate®) revealed similar transport behavior between the Transwell-based platforms. However, the OrganoPlate® exhibited significantly lower transport kinetics, compound movement being retarded by the presence of the gel channel. Finally, baseline gene expression analysis showed differences between cells cultured on Transwells and in 96-well plates or OrganoPlate®, however the presence of medium flow had minimal effects. Conclusions: This study demonstrates performance of different in vitro proximal tubule models with respect to their ability to address toxicity and pharmacokinetics. Based on gene expression data we found that culture platforms or presence of medium flow have relatively minor impacts on TERT-OAT1 RPTEC. Interestingly, the addition of fluid flow enhanced the sensitivity of nephrotoxicity detection but had little effect on drug transport. Our results provide clarity for optimizing platform selection for future studies of safety and pharmacokinetics and demonstrate the importance of carefully considering platform characteristics to balance physiological relevance, sensitivity, and throughput for specific contexts of use.