Abstract

Engineering of a Vascularized 3D Cell Construct On-Chip Using Human iPSC-derived Cells

Yu-suke Torisawa, Associate Professor, Hakubi Center for Advanced Research, Kyoto University

Vascular networks are essential to maintain cellular viability and function; however, current 3D culture models lack vascular systems.  Engineering perfusable vascular networks that can deliver reagents and blood cells to 3D cell constructs could be a powerful platform to recapitulate cellular microenvironments and tissue-level functions.  We have developed a microfluidic method to form vascularized tissue-like cell constructs to model cellular interactions through blood vessels.  When a tumor-like cell spheroid containing human umbilical vein endothelial cells (HUVECs) and fibroblasts was cultured in our microfluidic device, a perfusable vascular network was formed through the cancer spheroid.  We confirmed that peripheral blood mononuclear cells can be perfused inside a cancer spheroid through a vascular network.  Thus, we used this system to model the interaction between cancer cells and immune cells.  To study the interaction between cytotoxic T cells and cancer cells thorough blood vessels, allo-reaction between endothelial cells and T cells by mismatching of their HLA will be problematic.  Therefore, we engineered 3D vascular networks using human induced pluripotent stem cell-derived endothelial cells (hiPSC-ECs).  CD8+ T cells primed by HUVECs exhibited higher cytotoxic activity toward HUVECs than autologous hiPSC-ECs and MHC class I KO-hiPSC-ECs, demonstrating the potential value of this vascularized cancer-on-a-chip for modeling the interaction between T cells and a tumor-like tissue through blood vessels.  Generation of in vivo-like vascularized 3D cell constructs using hiPSC-ECs would provide a novel platform to develop organs-on-chips as well as human disease models.