Transparent Bioreactors Based on Nanoparticle-Coated Liquid Marbles for in Situ Observation of Suspending Embryonic Body Formation and Differentiation

Transparent liquid marbles coated with hydrophobic silica nanoparticles were used as micro-bioreactors for embryonic stem cell (ESC) culturing. The high transparency of silica liquid marbles enables real-time and in situ monitoring of embryonic body (EB) formation and differentiation. The experimental result shows that ESCs can aggregate with each other close to the bottom of the liquid marble and form EBs, while remaining suspended in the culture media. The differentiation of the suspending EBs into contractile cardiomyocytes has been demonstrated inside the transparent liquid marbles, which enable the in situ microscopic observation. It was also found, through comparison, that ESCs in a bare sessile drop placed on a superhydrophobic substrate tend to anchor onto the substrate and then differentiate following the normal way of cell spreading, i.e., withdrawal from the cell cycle, fusion with nascent myotubes, and final differentiation into cardiomyocytes. In contrast, liquid marble particle shells weaken the adhesion of spherical EBs to the substrate, encouraging them to differentiate in suspension into cardiomyocytes, without anchoring. The results of this study highlight the promising performance of liquid marbles as "one-pot" micro-bioreactors for EB formation and differentiation.