Abstract This project will commercialize a cell culture platform featuring SiMPore's ultrathin silicon- based membrane technology to enable advanced research on tissue barriers. In vitro models of tissue barriers such as the gut, lung, and vasculature are important for understanding the basis of disease and for assessing the ability of drug formulations to reach target tissues. Despite the growing use of sophisticated cell culture platforms (e.g., 3D cell culture, microphysiological systems, and tissue chips), the simplest and most popular tools for the in vitro study of barrier tissues remains the Corning Transwell™ and its competitors (collectively referred to herein as “Transwells™”). These products have a suspended ~ 10 µm thick polymer membrane creating apical and basal compartments which separate mono- or co-cultures grown on the membranes. Despite their popularity, Transwells® do not support high resolution microscopy nor provide the fluid flow needed to properly study vascular barriers and immune cell trafficking. SiMPore's membranes will be commercialized as cell culture products that overcome limitations of Transwells®, while retaining their easy to use format and offering features found in more sophisticated tissue chips. Our Phase I project successfully translated the laborious hand-made devices used in the laboratory of Professor James McGrath (University of Rochester) to a scalable fabrication workflow at SiMPore. Using a modular design, we developed an open-well Transwell-style culture unit that incorporates SiMPore's membranes, which further converts to a flow cell with the addition of a plug-and-play flow module. Devices were distributed to nine collaborating laboratories, all of whom reported success with the platform. SiMPore also successfully translated the dual-scale micro/nanoporous membranes developed by the McGrath laboratory to wafer-scale manufacturing. Our Phase II project will create commercially viable versions of Phase I prototypes to be marketed under the CytoVu™ brand. Aim 1 will increase membrane manufacturing capacity by relieving production bottlenecks and integrating automation. Aim 2 focuses on automating CytoVu™ device assembly. Aim 3 will test the manufactured devices in a network of collaborating laboratories while developing accessories that make the platform increasingly versatile and easy to use. This project will establish scalable manufacturing capacity at SiMPore for the CytoVu™ and its accessories, and also validate CytoVu™ products as competitive alternatives to incumbent products for the in vitro study of barrier tissues.