Innovative Cryopreservation Process Using a Modified Core/Shell Cell-Printing with a Microfluidic System for Cell-Laden Scaffolds

This work investigated the printability and applicability of a core/shell cell-printed scaffold for medium-term (for up to 20 days) cryopreservation and subsequent cultivation with acceptable cellular activities including cell viability. We developed an innovative cell-printing process supplemented with a microfluidic channel, a core/shell nozzle, and a low-temperature working stage to obtain a cell-laden 3D porous collagen scaffold for cryopreservation. The 3D porous biomedical scaffold consisted of core/shell struts with a cell-laden collagen-based bioink/dimethyl sulfoxide mixture in the core region and an alginate/poly­(ethylene oxide) mixture in the shell region. Following 2 weeks of cryopreservation, the cells (osteoblast-like cells or human adipose stem cells) in the scaffold showed good viability (over 90%), steady growth, and mineralization similar to those of a control scaffold fabricated using a conventional cell-printing process without cryopreservation. We believe that these results are attributable to the optimized fabrication processes the cells underwent, including safe freezing/thawing processes. On the basis of these results, this fabrication process has great potential for obtaining core/shell cell-laden collagen scaffolds for cryopreservation, which have various tissue engineering applications.