Architecture of an All‐In‐One Microfluidic Platform for Accelerated Cancer Seeding, Enhanced Spheroid Formation, and Dynamic Drug Screening Trials

ABSTRACT Effective cancer drug screening requires platforms that replicate the physiological complexity of tumor microenvironments. Traditional 2D systems fail to model critical features such as hypoxic gradients, interstitial flow, and drug penetration dynamics, resulting in limited predictive accuracy. To address these challenges, a microfluidic platform integrating porous V‐slanted hydrogel microwells, capable of forming uniform 3D cancer spheroids within the physiologically relevant size range of 200–500 µm, while enabling dynamic flow‐based drug testing is needed. This platform introduces advection‐based dynamic flow through a double‐layered microfluidic design, overcoming reliance on static diffusion seen in conventional systems. Dynamic drug screening performed on this platform demonstrates enhanced drug penetration and more consistent therapeutic responses compared to static conditions, emphasizing the importance of physiological flow in replicating in vivo tumor behavior. By controlling cross‐flow conditions, the platform ensures uniform drug delivery and enables a more reliable assessment of therapeutic efficacy, addressing variability often observed in static and 2D systems. Moreover, compatibility with high‐throughput applications makes it a scalable, robust solution for preclinical drug testing. This work highlights the critical role of dynamic environments in cancer drug screening, offering an improved physiologically relevant approach for studying drug responses and advancing personalized medicine strategies.