Abstract
The development of stable and standardized in vitro cytotoxicity testing models is essential for drug discovery and personalized medicine. Microfluidic technologies, recognized for their small size, reduced reagent consumption, and control over experimental variables, have gained considerable attention. However, challenges associated with external pumps, particularly inconsistencies between individual pumping systems, have limited the real-world application of cancer-on-a-chip technology. This study introduces a novel triplicate cell culture system (Tri-CS) that simultaneously supports dynamic cultures in three independent units using a single peristaltic pump, ensuring consistent flow conditions. Our findings demonstrate that the Tri-CS significantly reduces variability compared to individual pump systems, enhancing the reliability of anticancer drug cytotoxicity testing. Furthermore, we evaluated gemcitabine cytotoxicity, which shows enhanced drug efficacy in dynamic conditions. Fluorescein diffusion tests revealed greater diffusion efficiency in dynamic cultures, which contributed to the higher observed drug efficacy. The potential for broader application of the Tri-CS, including its compatibility with commercially available transwells and the opportunity for use in more complex cancer-on-chip models, positions this system as a valuable tool for advancing microphysiological systems in preclinical research.