Current organoid culture systems face critical limitations: standardized growth factor formulations fail to capture patient-specific signaling requirements, and single-cell-type approaches overlook tumor-stromal interactions essential for understanding immunotherapy resistance. To address these challenges, we developed an innovative biofabrication platform that systematically integrates patient-derived three-dimensional (3D) cultures with comprehensive growth factor profiling across 128 combinations. Through systematic screening of 23 ovarian cancer patient samples and single-cell RNA sequencing, we identified two estradiol-responsive cellular populations that coordinate immunosuppression: a malignant cell fraction (MAL.PDCD5) and a cancer-associated fibroblast (CAF) fraction (FB.TNFSF10). MAL.PDCD5 cells suppress immune infiltration by downregulating antigen presentation, while FB.TNFSF10 cells promote immunosuppression through enhanced TGF-β signaling. Spatial transcriptomic analysis revealed striking mutual exclusivity between FB.TNFSF10 cells and T/NK cells, providing evidence of active immune cell exclusion. Most significantly, FB.TNFSF10 abundance emerged as a robust predictor of immune checkpoint inhibitor therapy resistance across multiple cancer cohorts, independent of conventional biomarkers. This biofabrication platform provides a scalable framework for drug screening and biomarker discovery, with immediate applications in precision medicine for patient stratification and combination therapy development.