Modeling 3D Human Tumor Lymphatic Vessel Network Using High-Throughput Platform

The lymphatic vessel (LV) plays an important role in cancer biology as a major route for tumor metastasis. Whereas, recent oncoimmunological approaches have focused its role in immune surveillance. In response to the emerging topics of lymphatic vascular biology, physiologically relevant human cell-based in vitro model is in high demand. This study introduces a 3D in vitro model of human LV within tumor immune microenvironment (TIME) using an injection-molded plastic array culture platform (Lymph-IMPACT). Through spontaneous capillary flow-driven patterning of 3D cellular hydrogel and optimized cellular composition, the platform enables robust and reproducible formation of self-organized LV in vitro. This co-culture model recapitulates cancer cell type-dependent morphogenesis of LV in vitro. Moreover, the robustness of the model enables high-content analysis on the effect of anti-VEGFR3 drug depending on the existence of blood vessels or different types of cancer cells. By virtue of high perfusability of 3D lumenized in vitro LV, a trans-endothelial migration of cytotoxic primary lymphocytes, which is one of the critical processes in anti-tumor immunology, is recapitulated within reconstructed melanoma TIME. From drug testing to cellular migration assays using the high-throughput platform, the Lymph-IMPACT demonstrates its powerful potential to be applied on investigating lymphatic-related strategies for cancer therapeutics.