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

Abstract

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.

On‐Demand Coalescence and Splitting of Liquid Marbles and Their Bioapplications

“…In a recent publication, the researchers suggested that the liquid marble-based 3D stem cell culture showed its potential applications in embryonic body formation and differentiation. [46] The viability of cells growing inside the LMs was determined using the 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. As shown in Figure 7e, the optical density (O.D.)…”

“…The 3D SCSs that self‐assembled at a density of 10 6 cells mL −1 was tightly packed with ≈5000 cells in each spheroid, creating an “in vivo‐like” micro‐bioenvironment for organoid development that better preserved the stem cell phenotype and its innate properties. In a recent publication, the researchers suggested that the liquid marble–based 3D stem cell culture showed its potential applications in embryonic body formation and differentiation …”

Liquid Marbles as Miniature Reactors for Chemical and Biological Applications

“…The capability of introducing reagents and withdrawing products is crucial for liquid marbles to operate effectively as microreactors. Previous studies have reported the use of micropipettes to add and extract interior liquids without damaging liquid marbles [43][44][45][46]. However, this method requires the micropipette tips to penetrate the coating shell of liquid marbles, making the interior liquids susceptible to contamination.…”

“…The embryoid bodies differentiated into contractile cardiomyocytes, which can be used in cell replacement therapy. Lin et al utilised transparent liquid marbles covered with silica nanoparticles for the growth of embryoid bodies from Oct4B2-ESC and their differentiation into cardiomyocytes without transferring the embryoid bodies into another culture platform (Figure 9b) [45]. The transparency of liquid marbles allows for continuous and real-time monitoring of the formation of embryoid bodies and, eventually, the differentiation into contractile cardiac cells.…”

Urea Melt Marbles Developed by Enwrapping Urea Melt Droplets with Superhydrophobic Particles: Preparation, Properties, and Application in Large Urea Granule Production

“…[1] However, the main industrial technologies used for urea granulation, including a prilling tower and fluidized bed granulation, have difficulty adhering to such limits owing to the insuperable dust generation according to the granulation principle, and use of an expensive and energy this range, the outer hydrophobic particle layer prevents the destruction or coalescence of liquid marbles, protecting the internal droplets from becoming independent entities. [9] Owing to their special properties, liquid marbles have been employed in various interesting applications, including those in microreactors, [10] capsule preparing, [11] microbial culture media, [12] microfluid delivery, [13] electrical sensor, [14] and magnetic sensors. [15] The merits of liquid marbles are exactly in line with the required demands of the DLUG process for continuous operation.…”