Fig. 1: Conceptual 2D representation and workflow of the 96-wells fluidic system.

Step 1: The 96-islands PDMS substrate is obtained through PDMS molding by using the PMMA 96-wells master. Step 2: The 96-islands PDMS substrate and the 96-well cell culture part (made of PMMA) are sterilized and assembled by inserting the 96-islands PDMS substrate into the 96-well cell culture part. The parts are bond together by using commercial metal binder 1.6 inches wide paper clamps on the sides. This creates the 96-wells cell culture component. Step 3: Each well of the 96-islands PDMS substrate is coated with the ECM of interest followed by seeding of cells on each PDMS well. Step 4: Adherent cells will grow forming a monolayer. A lid of a regular cell culture plate is used in order to maintain aseptic cell culture conditions. At this step, cell manipulation can be performed, such as siRNA transfection. Cell monolayers will remain in the 96-well cell culture component until ready to be exposed to flow. Step 5: The 96-wells cell culture component, with the cells, is disassembled by removing the lid and the 96-well culture part. The flow channels part, containing the inlet/outlet and channels part, is then inserted on top of the 96-islands PDMS substrate and the PMMA base to create the 96-wells flow component. The channels part contains 96 cavities of the same height as the PDMS islands. When assembled, the 96-wells flow component creates 8 linear and independent flow channels of 500 µm height. Step 6: The 96-wells flow component is ready to be coupled to a fluidic system via elbow luer connectors, or equivalent, allowing perfusion of cell culture media at the desired flow rate