Horseradish peroxidase-mediated encapsulation of mammalian cells in hydrogel particles by dropping

We report a method for preparation of mammalian cell-enclosing hydrogel particles through horseradish peroxidase (HRP)-catalysed hydrogelation by dropping cell-suspending aqueous solution into an aqueous coagulation solution. An aqueous solution of 10% (w/v) gelatin derivative possessing phenolic hydroxyl (Ph) moieties (Gelatin-Ph), HepG2 cells and 10 U/mL HRP was dropped into an aqueous coagulation solution containing 1 mM H2O2. The resultant hydrogel formed through the HRP-catalysed reaction consuming H2O2 had a spherical shape. The sphericity decreased with decreasing concentrations of Gelatin-Ph, HRP and H2O2. The thickness of the hydrogel membrane layer of the hydrogel particles could be controlled by altering incubation time in the H2O2 solution. The cells encapsulated in the particles with a thinner hydrogel membrane grew faster. These results demonstrate that we successfully established the method of cell-encapsulation in hydrogel particles based on dropping aqueous polymer solution into aqueous coagulation solution through HRP-catalysed reaction.