Optimized cell mixing facilitates the reproducible bioprinting of constructs with high cell viability

Abstract

Bioprinting with cell-laden hydrogels (bioink) requires the careful mixing of cells with the hydrogel carrier to ensure that the bioink is homogeneous and functional, and the printing results are reproducible. Bioink preparation is therefore a critical process step that must accommodate the specific rheological properties of different bioinks. Here, we developed a reproducible method for the optimized mixing of cells and hydrogel carriers that can be integrated into current bioprinting processes. First, we tested and optimized different mixing devices for their effect on bioink homogeneity and rheological properties, resulting in a low-shear process for the preparation of homogenous bioinks. Based on these findings, we evaluated the impact of different cell densities on the rheological profile of bioinks according to shear and temperature, and estimated the impact of shear stress intensity and duration on 1.1B4 cells. Finally, we integrated the optimized mixing method into a current printing process and monitored the printed construct for 14 days to confirm cell viability. We found that the cell viability in the printed cell-laden constructs remained in excess of 91% after 14 days.