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

Abstract

Abstract 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.