Superhydrophobic Chips for Cell Spheroids High-Throughput Generation and Drug Screening

We suggest the use of biomimetic superhydrophobic patterned chips produced by a benchtop methodology as low-cost and waste-free platforms for the production of arrays of cell spheroids/microtissues by the hanging drop methodology. Cell spheroids have a wide range of applications in biotechnology fields. For drug screening, they allow studying 3D models in structures resembling real living tissues/tumors. In tissue engineering, they are suggested as building blocks of bottom-up fabricated tissues. We used the wettability contrast of the chips to fix cell suspension droplets in the wettable regions and evaluated on-chip drug screening in 3D environment. Cell suspensions were patterned in the wettable spots by three distinct methods: (1) by pipetting the cell suspension directly in each individual spot, (2) by the continuous dragging of a cell suspension on the chip, and (3) by dipping the whole chip in a cell suspension. These methods allowed working with distinct throughputs and degrees of precision. The platforms were robust, and we were able to have static or dynamic environments in each droplet. The access to cell culture media for exchange or addition/removal of components was versatile and opened the possibility of using each spot of the chip as a mini-bioreactor. The platforms' design allowed for samples visualization and high-content image-based analysis on-chip. The combinatorial analysis capability of this technology was validated by following the effect of doxorubicin at different concentrations on spheroids formed using L929 and SaOs-2 cells.