High-throughput fabrication of HAMA/matrigel core-shell hydrogel microspheres by droplet microfluidics for in vitro NSCLC model

Three-dimensional (3D) in vitro tumor models are increasingly used to better recapitulate key structural features of solid tumors than conventional two-dimensional culture systems. However, the high-throughput fabrication of uniform 3D tumor constructs remains challenging. Here, we report a droplet microfluidic platform for the high-throughput generation of tumor-laden core-shell microspheres. The platform consists of a three-channel microfluidic device, with Matrigel as the core phase and HAMA as the shell phase. Through coaxial flow and oil-phase shearing, core-shell droplets were generated and subsequently stabilized by blue-light photocrosslinking of the HAMA shell. By adjusting the flow rates of the core, shell, and oil phases, we obtained core-shell microspheres with controllable size and good uniformity. Under optimized conditions, including core and shell flow rates of 10:10 µL/min, an oil flow rate of 200 µL/min, and 4% HAMA, the platform generated microspheres with diameters of approximately 450 μm and enabled the construction of an in vitro NSCLC microsphere model.We then performed an initial characterization of the resulting constructs, including morphology, cell viability, and response to a model anticancer drug. Compared with a conventional 3D culture approach, the core-shell microspheres showed improved structural integrity and supported cell survival over the tested culture period. These findings demonstrate the feasibility of this microfluidic fabrication strategy for producing tumor-laden core-shell microspheres and suggest its potential as a basis for future in vitro tumor modeling and drug-response studies.