Vertical Membrane‐Free Organ‐on‐a‐Chip for High‐Throughput In Vitro Studies and Drug Screening

Abstract To bring in vitro models to a reliable and widely adopted experimental platform for preclinical and therapeutic research, we introduce a scalable, and parallelized microfluidic plate for Organ‐on‐a‐Chip (OoC) applications. This OoC unit consists of two vertically interconnected channels with a highly hydrophilic base layer which allows the flow within the bottom channel without invading the top channel. Vertical cell compartmentalization without the use of a porous membrane or physical obstacles enables perpendicular screening and semicircular cell organization for homogeneous shear stress. Fabricated using standard materials and techniques, it is accessible to a wider range of users. The plate contains three‐layered design: a transparent poly(methyl methacrylate) (PMMA) microfluidic chip, a medical‐grade pressure‐sensitive adhesive layer, and a hydrophilic sealing layer. The OoC unit was parallelized into a standard 384‐well plate format, enabling high‐throughput use. We demonstrated its versatility by successfully generating models of intestine and musculoskeletal tissue models, and validated for various applications, including drug discovery, personalized medicine, and disease modeling. Overall, this novel OoC unit has demonstrated the potential to predict different physiological features and can be used for pharmacological and toxicological studies, while decreasing the need and use of animals required during drug development.