3D-Cell Culturing Chambers(CCC) integrated Drug Screening Platform with Linear & Logarithmic Concentration Gradients for Anti-Cancer Drug

Abstract

A double-layered platform, with a concentration gradient generator (CGG) and 3D cell culturing chambers (CCC), was designed to generate 10 different concentration levels of linear and logarithmic(3-order) gradients. Hagen-Poiseuille flow resistance equation and the parallel electric schematics were used to calculate the channel dimensions for 100%, 98%, 50%, 30%, 20%, 10%, 5%, 2%, 1%, and 0% drug concentration. The designated concentration levels were compared with the predicted values by running fluorescein dye and PBS in place of drug and diluting agent, respectively. T24 cells (human bladder cancer cells) embedded in the fibrin gels were loaded and monitored on the platform for 72 hours. At the end of the 3-day experiment, live-dead reagents and Presto Blue reagent were injected directly into the chip to evaluate the cell viability. The culturing chambers were laid out at an interval of 9mm to match the well-plate spacing for a microplate reader. The platform’s vertical layout minimizes the lateral dimension required for housing various components while maintaining a favorable height for imaging. By preventing the need to use external tubing to connect the two (CGG and CCC) separate entities, this platform will facilitate transitioning to screen combinatory therapy and other drug evaluations.