Cell-laden hydrogel microcarriers are widely used in diverse biomedical applications like three-dimensional (3D) cell culture, cellular therapy, and tissue engineering, where microcarriers were generally produced by oil, which is the common but not optimal choice, as oil may cause cytotoxicity or protein denaturation. Here, an all-aqueous-phase microfluidics is presented to achieve oil-free emulsification of cell-laden microcapsules and 3D cell culture. Aqueous solutions with different concentration gradients are used as an immiscible continuous phase and a dispersed phase, and oscillation from a solenoid valve facilitates the formation of microcapsules at the water–water interface. By adjusting aqueous-phase flow rates and oscillating frequencies, core–shell microcapsules with controllable structures can be stably and continuously generated. In further 3D cell culture, encapsulated cells maintained good viabilities and aggregated together. These features show that the oil-free microfluidic method may have broad prospects in many biomedical applications.

Core-shell structured stem cell microencapsulation in hydrogel has wide applications in tissue engineering, regenerative medicine, and cell-based therapies because it offers an ideal immunoisolative microenvironment for cell delivery and 3D culture and differentiation. Long-term storage of such microcapsules as cell-biomaterial constructs by cryopreservation is an enabling technology for their wide distribution and ready availability for clinical transplantation. However, most of the existing studies focused on cryopreservation of separated cells or cells in microcapsules without a core-shell structure (i.e., hydrogel beads). The goal of this study is to achieve cryopreservation of stem cells encapsulated in core-shell microcapsules as cell-biomaterial constructs or biocomposites. To this end, a capillary microfluidics-based core-shell alginate hydrogel encapsulation technology is developed to facilitate cryopreservation of porcine adipose-derived stem cells (pADSCs) laden microcapsules with very low concentration (2 mol L−1) of cell membrane penetrating cryoprotective agents (CPAs) by suppressing ice formation. This may provide a low-CPA and cost-effective approach for vitreous cryopreservation of “ready-to-use” stem cell-biomaterial constructs, facilitating their off-the-shelf availability and widespread applications.

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