3D-Printed Microfluidic-Based Cell Culture System With Analysis to Investigate Macrophage Activation

In this paper, we describe the development of 3D-printed microfluidic cell culture devices that can be coupled with a circulation system to study the dynamics of both intracellular and extracellular (release) processes. Key to this approach is the ability to quantitate key analytes on a minutes timescale with either on-line (in this study, quantitating nitric oxide production using an amperometric flow cell) or off-line (in this work, quantitating intracellular itaconate production with LC/MS) analytical measurements. To demonstrate the usefulness of this approach, we chose to study macrophage polarization as a function of the extracellular matrix (silk) fiber size, a major area of research in tissue engineering. It was found that the use of larger fibers (1280 nm vs. smaller 512 nm fibers) led to increases in the production of both nitric oxide and itaconate. These findings set the foundation for future research for the creation of finely tuned microfluidic 3D cell culture approaches in areas where flow and the extracellular matrix play a significant role in barrier transport and where integrated analysis of key markers is needed.