Microwell arrays for uniform-sized embryoid body-mediated endothelial cell differentiation

Embryonic stem (ES) cell is of great interest cell source in regenerating tissue constructs. We hypothesized that the interaction of cell-extracellular matrices (ECMs) would enable the control of ES cell differentiation pathway. We fabricated the hydrogel microwell array system to regulate uniform-sized embryoid bodies (EBs) and replate into various ECM components (e.g., gelatin, collagen I, fibronectin, laminin, and Matrigel). We demonstrated that collagen I and laminin largely induced ES cell-derived endothelial cell differentiation compared to gelatin. We also characterized ECMs-dependent endothelial cell differentiation by evaluating the endothelial gene expression, showing that Flk1 endothelial gene was highly expressed on collagen I. We also demonstrated the effect of the integrin on uniform-sized EBs-derived endothelial cell differentiation, showing that integrin α1 was largely expressed on laminin. Therefore, the cell-ECM interaction could be potentially powerful for controlling the uniform-sized EBs-derived endothelial cell differentiation.