High-Throughput 3D Spheroid Production with Photocurable Polyurethane Microwell Arrays via LCD 3D Printing

A photocurable and biocompatible polyurethane resin was synthesized and processed via liquid crystal display (LCD) 3D printing to produce high-resolution elastomeric microwell arrays for spheroid generation. The resin was characterized by a low viscosity of 384 mPa.s and an elastic modulus of 2.7 ± 0.2 MPa post-printing. Utilizing an XY resolution of 22 µm and 10 µm layer thickness, the resulting customizable devices featured non-adhesive, hydrophilic surfaces that promoted cell-cell interactions over substrate attachment. Validation using human lung adenocarcinoma (A549) cells confirmed the formation of uniform spheroids with high viability (84 ± 6%) and no evidence of a necrotic core after 7 days of culture. Quantitative morphological analysis revealed high structural regularity, with roundness values between 0.83 and 0.90 and solidity consistently exceeding 0.92. Proliferation within the device was further supported by a time-dependent increase in DNA content. This additive manufacturing approach offers a precise, high-resolution, and customizable alternative to conventional molding for the fabrication of 3D cell culture platforms, with significant applications in in vitro disease modeling, stem cell delivery, and organoid research.