Hydrogel Encapsulation Facilitates Rapid-Cooling Cryopreservation of Stem Cell-Laden Core–Shell Microcapsules as Cell–Biomaterial Constructs

Abstract

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. 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 focus on cryopreservation of single 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 produce porcine adipose-derived stem cell-laden microcapsules for vitreous cryopreservation 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.

Alginate hydrogel encapsulation by capillary microfluidics facilitates rapid-cooling cryopreservation of stem cell-laden core–shell microcapsules as cell–biomaterial constructs. It enables vitreous cryopreservation of encapsulated stem cells with very low concentrations of penetrating cryoprotective agents, providing a cost-effective and high-throughput approach for long-term storage of stem cells and stem cell–biomaterial constructs.

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