An accessible and open-source workflow for fabrication of custom PDMS culture chambers using 3D printing.

Custom cell culture platforms are widely used in tissue engineering applications. However, their fabrication often requires specialized microfabrication equipment and advanced technical skills, limiting accessibility. This work presents an accessible and open-source workflow for the fabrication of custom PDMS culture chambers. This workflow focuses on the use of parametric CAD designs, 3D-printed molds, resin casting and plasma-free bonding. The proposed approach allows users to generate customized molds adapted to the experimental requirements using a graphical interface without extensive CAD knowledge. The molds can be manufactured using commonly available 3D printers or outsourced at relatively low cost. Device assembly is performed using uncured PDMS as a bonding agent, eliminating the need for plasma bonding equipment. The resulting culture chambers were validated through watertightness testing and biological evaluation using N2a cell cultures. Cell viability after 72 h of culture was comparable to that observed in standard culture plates. By reducing requirements for specialized equipment and technical expertise, this workflow facilitates the rapid, low-cost and reproducible fabrication of customized PDMS culture platforms for tissue engineering and related biomedical applications.