Engineering a Functional Angiogenesis-Enabled and Ovarian Cell-Specific Tumor Microenvironment Chip

Abstract Purpose Angiogenesis plays a critical role in ovarian cancer progression, yet physiologically relevant in vitro models that recapitulate ovarian cell-specific tumor-vascular interactions remain limited. Standardized workflows for ovarian cancer spheroid generation, vascularization, and functional angiogenesis assessment within microphysiological systems are also lacking. Here, we developed a vascularized ovarian tumor microenvironment (TME)-on-chip platform and established a standardized modular framework for ovarian cancer modeling and angiogenesis studies. Methods We systematically compared ovarian cancer cell lines and spheroid formation methods to define reproducible tumor modules with distinct morphological stability and pro-angiogenic phenotypes. Following on-chip integration, tumor viability, structural stability, and invasive behavior were evaluated. Multiple endothelial cell sources were compared to establish an ovarian cell-specific vascular component. Disease-relevant gain- and loss-of-function studies were subsequently performed using lysophosphatidic acid (LPA) and Bevacizumab, respectively. Results On-chip tumoroids maintained viability, structural stability, and invasive behavior throughout culture. Human ovarian microvascular endothelial cells (HOMECs) supported robust lumen formation, angiogenic remodeling, and tumor-associated vascular interactions. LPA treatment enhanced VEGF secretion and angiogenic network formation, whereas anti-VEGF treatment with Bevacizumab suppressed tumor-driven angiogenesis. Conclusions This study establishes a standardized, biologically responsive, and ovarian cell-specific vascularized TME-on-chip model for investigating tumor-microvascular crosstalk, angiogenic signaling, and anti-angiogenic therapeutic responses, with future potential for incorporating stromal and immune components.