“Cell-Capture Pitfall” by Two-Layered Microwell Array Device for Single-Cell Derived Exosome Analysis

This paper describes a two-layered hydrogel microwell array device for single-cell-derived exosome analysis with a cell capture system named “Cell-capture pitfall”. Our proposed device consists of (i) a first layer of a microwell array with a soluble bottom surface and (ii) a second layer of cell-culture hydrogel array. This device enables efficient single cell capture and long-term culture while isolating exosomes. We achieved a cell capture efficiency exceeding 70% and confirmed that the cells transfer from the upper to the lower wells using the “Pitfall method”. Furthermore, we maintained viable cell cultures in the device for over 17 days. We believe that this device offers a powerful tool for single-cell exosome analysis and holds promise for advancing our understanding of cancer pathophysiology.