High‐Throughput Microspheres Production Assisted by Hydrophobic Interface for Tumor Organoid Culture and Drug Evaluation

Abstract

Conventional microfluidic droplet techniques requiring a long output pipeline to ensure the gelation of Matrigel microspheres, which are often compromised by droplet coalescence, tubing clogging, and poor system stability. To overcome these limitations, a well‐plate‐based in situ gelation strategy was developed. This approach relocated the gelation process from lengthy tubing to a receiving well‐plate deposited with hydrophobic particles, thereby enabling short‐path droplet transport and off‐tubing gelation. The hydrophobic interface at the well bottom was systematically screened and optimized. Silanized silica nanoparticles were identified as an effective modifier. This layer maintained high optical transparency and demonstrated favorable biocompatibility while effectively preserving the spherical morphology of the droplets. Matrigel microspheres encapsulating A549 cells and patient‐derived lung cancer organoids were successfully fabricated using this system. Chemosensitivity testing demonstrated that the prepared microspheres exhibited differential responses to chemotherapeutic agents and radiotherapy across the various models, comparable to those from traditional long‐tubing preparation systems. The integrated platform provides a reliable technical solution for high‐throughput, standardized tumor organoid culture and personalized therapeutic screening.