A Simple Pipetting-based Method for Encapsulating Live Cells into Multi-layered Hydrogel Droplets

Abstract

Here, we present a simple pipetting-based approach for generating multi-layered core-shell hydrogel droplets composed of alginate for outer shell and collagen for culturing cells inside. By using a multi-hole plastic substrate and by pipetting, multi-layered hydrogel droplets were generated in a simple and rapid manner. HEK293 cells, which are human embryonic kidney cells, were cultured for 14 days in double-layered hydrogel droplet with high viability. Cancer cells were co-cultured with epithelial cells in multi-layered hydrogel droplets and applied for drug tests with curcumin. As epithelial cells protect cancer cells from anti-cancer drugs, co-cultured cells showed lower sensitivity to curcumin. We developed a simple and easy method for creating complex hydrogel particles for 3D multicellular co-culture and developed an alternative method for drug testing in vivo.