High-Throughput Digital Decoding of Vascular Heterogeneity in Patient-Specific Tumor Microenvironments.

Quantitative characterization of vascular heterogeneity in complex microphysiological systems (MPS), particularly within patient-derived tumor microenvironments, remains a major challenge for scalable disease modeling and therapeutic evaluation. Existing analysis approaches primarily rely on vessel abundance-based morphometrics and often fail to resolve network connectivity and spatial remodeling in heterogeneous vascular systems. Here, we present the iMAP platform, a high-throughput digital vascular profiling framework that integrates an injection-molded microfluidic chip with an interactive image analysis tool (iMAP Analyzer). This platform enables topology-resolved and region-aware quantification of vascular architecture, capturing vessel morphology, branching complexity, connectivity, and spatial variation between tumor-proximal and distal regions. Applied to 3D co-cultures of patient-derived gastric cancer tumor spheroids with either donor-matched iPSC-derived endothelial cells (iPSC-ECs) or primary HUVECs, iMAP identified differences in network organization and spatial stability under standardized conditions, with iPSC-derived networks exhibiting increased fragmentation and peripheral instability. These findings demonstrate that connectivity-normalized and region-resolved analysis provides critical insight beyond conventional whole-image metrics. The iMAP framework offers a scalable and standardized approach for quantitative vascular phenotyping in complex MPS, supporting high-content analysis and advancing the development of vascularized in vitro disease models.