Gut-on-Chip Methodology Based on 3D-Printed Molds: A Cost-Effective and Accessible Approach

Abstract

1Gut-on-chips (GoC) represent a disruptive technology with great potential to understand the underlying mechanisms of gut health and pathology. Researchers who want to implement this approach, can either use expensive commercial microfluidic chips or build them from scratch in their lab. However, the design of such devices demands specific technical skills and expertise in computer assisted design (CAD). Additionally, the fabrication of the master molds for the chip production is very costly, time consuming and requires dedicated microfabrication facilities. Thus, the diffusion of these models in biology and health research laboratories remains limited due to this technological complexity and lack of affordability. In order to break these two bottlenecks, we present here the 3DP-{micro}Gut with open access designs and a simple fabrication approach based on a commercial 3D printer intended for general users. To ensure a good reproducibility with a sufficient number of available replicates for biological experiments, the method has been optimized to allow the production of multiple GoC per batch. The chips were also improved for confocal live imaging microscopy analyses. The chips are designed to be compatible with different types of microfluidics pumps, from stand-alone to completely integrated instrumentation. As a proof of concept, Caco-2 cells were seeded inside the fabricated GoC to validate their biocompatibility and functionality. After 7 days of maturation, cells self-differentiated in a 3D epithelium resembling in vivo expected structures. In summary we proposed a low-cost and open access GoC design and fabrication method with medium throughput. These results demonstrate the advantage of using 3D SLA printing to accelerate GoC implementation for gut physiopathology research.