Storable Cell-Laden Alginate Based Bioinks for 3D Biofabrication

Over the last decade, progress in three dimensional (3D) bioprinting has advanced considerably. The ability to fabricate complex 3D structures containing live cells for drug discovery and tissue engineering has huge potential. To realise successful clinical translation, biologistics need to be considered. Refinements in the storage and transportation process from sites of manufacture to the clinic will enhance the success of future clinical translation. One of the most important components for successful 3D printing is the 'bioink', the cell-laden biomaterial used to create the printed structure. Hydrogels are favoured bioinks used in extrusion-based bioprinting. Alginate, a natural biopolymer, has been widely used due to its biocompatibility, tunable properties, rapid gelation, low cost, and easy modification to direct cell behaviour. Alginate has previously demonstrated the ability to preserve cell viability and function during controlled room temperature (CRT) storage and shipment. The novelty of this research lies in the development of a simple and cost-effective hermetic system whereby alginate-encapsulated cells can be stored at CRT before being reformulated into an extrudable bioink for on-demand 3D bioprinting of cell-laden constructs. To our knowledge the use of the same biomaterial (alginate) for storage and on-demand 3D bio-printing of cells has not been previously investigated. A straightforward four-step process was used where crosslinked alginate containing human adipose-derived stem cells was stored at CRT before degelation and subsequent mixing with a second alginate. The printability of the resulting bioink, using an extrusion-based bioprinter, was found to be dependent upon the concentration of the second alginate, with 4 and 5% (w/v) being optimal. Following storage at 15 °C for one week, alginate-encapsulated human adipose-derived stem cells exhibited a high viable cell recovery of 88 ± 18%. Stored cells subsequently printed within 3D lattice constructs, exhibited excellent post-print viability and even distribution. This represents a simple, adaptable method by which room temperature storage and biofabrication can be integrated for on-demand bioprinting.