Low-Cost,Rapid Fabrication of Customizable Polyethylene Glycol-Based Cell Culture Devices

Abstract

Biological research groups may face a high barrier to entry when constructing custom 3D cell culture devices to investigate multi-tissue interactions in vitro. Standard fabrication methods such as lithography, etching, or molding are expensive and require specialized equipment and expertise. To address this, we developed an accessible approach for producing polyethylene glycol (PEG)-based cell culture devices using stereolithography 3D printing with a polydimethylsiloxane intermediate mold. Both the intermediate molding steps and the sterilized final device show low cytotoxicity, and the final device swells to predictable dimensions and retains its shape for at least 10 days. We used this approach to develop a human pluripotent stem cell-derived neural spheroid outgrowth model that supports directed neurite extension over 14 days. Together, this method provides a highly customizable, affordable platform for rapid fabrication of PEG-based microphysiological systems for diverse tissue engineering applications.

Impact Statement

As biomedical labs work to complement animal models with tissue-engineered microphysiological systems (MPSs), there is a growing need for low-cost, rapid, and iterative fabrication workflows. We developed a pipeline combining 3D printing, a polydimethylsiloxane intermediate mold, and polyethylene glycol casting, avoiding the need for specialized photolithography. The resulting devices support stable, nutrient-permissive cell culture while allowing control over device dimensions and customizable channel or compartment configurations. We demonstrate the device’s utility using reprogrammed human pluripotent stem cell-derived neurons, which remain challenging to support sustained neurite outgrowth in engineered models. This workflow expands access to cell culture device fabrication for MPSs across a broader range of biological laboratories.