The Texas A&M Tissue Chip Validation (TEX-VAL) Consortium is a public-private partnership that aims to promote the use of microphysiological systems (MPS). The Consortium is engaged in stakeholder needs-directed applied research to establish the functionality, reproducibility, robustness, and reliability of a wide array of MPS. One high-throughput MPS for liver studies was recently proposed in which induced pluripotent stem cell-derived hepatocytes (iHeps) and non-parenchymal cells (NPCs) were co-cultured in the OrganoPlate 2-lane plate containing 96 independent MPS. Our goal was to evaluate this platform using additional cell types and conditions and characterize its utility and reproducibility. Primary human hepatocytes (PHHs) or iHeps, with and without NPCs (vascular endothelial and THP-1 monocytes), were cultured for up to 17 days. Imaging-based cell viability, albumin and urea secretion into culture media, and CYP3A4 activity were assessed using midazolam, a CYP3A4 substrate. The iHeps co-cultured with NPCs formed cell aggregates in the gel lane and demonstrated stable cell viability and function up to 17 days in culture in most of the experiments; however, variability was appreciable across replicates and replicate studies. For iHeps monoculture, cells did not form clusters and lost viability and function over time. For PHH monoculture, cell viability and hepatic function were also low and not sustained over time. The disappearance of midazolam and the formation of its metabolites were most efficient under the iHeps and NPCs co-culture conditions. Overall, we find that this platform used with iHeps and NPCs is a functional liver MPS model that can be used to assess metabolism; however, the high-throughput nature of this MPS is somewhat dampened by the need for replicates of each condition to compensate for high variability. This abstract does not represent policy or product endorsement by TEX-VAL Consortium member organizations American Chemistry Council, BristolMyers Squibb, Merck KGaA, National Institute of Environmental Health Sciences, Sanofi-Aventis, Unilever, and US EPA.