3D Printed Microfluidic-Based Cell Culture System with Analysis to Investigate Cell-to-Cell Communication

Most cell culture studies are in 2D, which may not accurately represent the cellular microenvironment in vivo, where many cells grow in a 3D fashion within the network of extracellular matrix (ECM). To mimic ECM features, we electrospun polystyrene (PS) scaffolds and incorporated them into 3D-printed insert-based devices for cell culture. Inset-based devices allow different cell types to be cultured on nanofiber scaffolds, which can be easily placed and removed from the device for analysis.
In this study, PolyJet and Fused Deposition Modeling (FDM) printing techniques were used to create insert-based devices and petri dish-like devices for cell culture under static and flow conditions, respectively. These printer types were employed to evaluate how their printing materials affect cell viability. Unlike PolyJet-printed devices, FDM-printed devices usually leak between layers. We have optimized parameters that allow the printing of leak-free FDM devices using PS material, a polymer commonly used to make cell culture flasks. These devices withstood leakage when subjected to flow experiments. Furthermore, the devices have been optimized by reducing dead volume, improving cell immobilization, and assessing cell viability for long-term cell culture. NMR spectroscopy has been utilized to investigate leachates of PS material resulting from the FDM printed devices.
Bovine pulmonary artery endothelial cells were used in this study because of their role in regulating vasodilation by releasing nitric oxide (NO) in response to shear stress or ATP stimulation. Red blood cells (RBCs) can also release both ATP and NO when they undergo mechanical deformation. Consequently, these cells can communicate by releasing these molecules. We have used PolyJet printing to create a multimodal device that allows us to simultaneously detect NO (via amperometric) and ATP (via chemiluminescence) from RBCs. We will present this, as well as a study to measure NO release from endothelial cells.