Regenerating the cell resistance of micromolded PEG hydrogels

Polydimethylsiloxane stamp materials used during soft lithography undermine the non-fouling behaviour of bio-inert PEG-based hydrogels, resulting in increased protein adsorption and cell adhesion and migration on the gel. This previously unreported phenomenon undermines the function of lab-on-a-chip devices that require the device to be bio-inert, and slows the implementation of promising micromolding and imprinting methods for 3D culture and commercial cell culture systems. We illustrate that the degree of cell adhesion and protein adsorption to the gels correlates with the amount of residual stamp material remaining at the hydrogel interface after fabrication. After identifying this previously unreported phenomenon, we screened multiple polymer cleaning/fabrication techniques in order to maintain/restore the non-fouling properties of the gels including PDMS curing and extraction, use of other common soft lithography stamp materials, post-fabrication cleaning of the hydrogels, and changing the composition of the hydrogel. The optimal solution was determined to be incorporation of reactive sites into the hydrogel during micromolding followed by grafting of PEG macromers to these sites post-fabrication. This treatment resulted in micromolded hydrogels with robust cell resistant properties. Broadly, this work identifies and solves a previously unreported problem in hydrogel micromolding, and specifically reports the development of a cell culture platform that when combined with video microscopy enables high-resolution in situ study of single cell behaviour during in vitro culture.