Customized 3D-printed stackable cell culture inserts tailored with bioactive membranes.

Area of Science:

• Biotechnology
• Materials Science
• Cell Biology

Background:

• Transwell inserts are widely used for cell culture but lack customization options.
• Customization of cell culture inserts can improve experimental outcomes for complex models.

Purpose of the Study:

• To develop customizable 3D-printed cell culture inserts with functionalized membranes.
• To demonstrate the feasibility of on-demand printing and functionalization for diverse laboratory needs.

Main Methods:

• Fabrication of a library of 3D-printed inserts using stereolithography (SLA).
• Functionalization of insert apertures with hydrogels via a high-throughput dip-casting technique.
• Comparison of mouse intestinal organoid development on different insert bottom materials.

Main Results:

• Successfully created 3D-printed inserts in various formats (24- to 96-well).
• Achieved well-defined hydrogel membranes with low variability (<10% CV) and thickness of ~500 µm.
• Demonstrated consistent intestine barrier formation over 3 weeks and distinct organoid condensation patterns based on insert bottom type.

Conclusions:

• 3D-printed cell culture inserts offer a customizable and realistic alternative to standard devices.
• The choice of insert bottom material (hydrogel vs. synthetic filter) is critical and application-dependent.
• On-demand printing of tailored cell culture inserts is feasible for research laboratories.