Fabrication of hexagonally packed cell culture substrates using droplet formation in a T-shaped microfluidic junction

A method is here proposed to fabricate ordered hexagonally packed cell culture substrates with hexagonally arranged cell patterning areas. We generated photo-sensitive polymeric microdroplets in a T-shaped microfluidic junction by an immiscible liquid, and then solidified the collective self-assembled hexagonal droplet array to obtain the cell culture substrate, on which we took the grooves formed between the solidified droplets as the hexagonally arranged cell patterning areas. The most promising advantage of our method is that we can actively tune the droplet size by simply adopting different volumetric flow rates of the two immiscible fluids to form cell culture substrates with differently sized cell patterning areas. Besides, the examination results of the cell culture substrate's characteristics validate whether our method is capable of creating substrates with high spatial uniformity. To verify the cell patterning function of our cell culture substrates, we used the semi-adherent RAW cells to demonstrate the effectiveness of patterning of suspended/adherent cells before/after adhesion. Over 90% cell viability and cell patterning rate suggest that our method may be a promising approach for future applications of cell patterning on biochips.