A novel cell-printing method and its application to hepatogenic differentiation of human adipose stem cell-embedded mesh structures

摘要

We report a cell-dispensing technique, using a core–shell nozzle and an absorbent dispensing stage to form cell-embedded struts. In the shell of the nozzle, a cross-linking agent flowed continuously onto the surface of the dispensed bioink in the core nozzle, so that the bioink struts were rapidly gelled, and any remnant cross-linking solution during the process was rapidly absorbed into the working stage, resulting in high cell-viability in the bioink strut and stable formation of a three-dimensional mesh structure. The cell-printing conditions were optimized by manipulating the process conditions to obtain high mechanical stability and high cell viability. The cell density was 1?×?107?mL?1, which was achieved using a 3-wt% solution of alginate in phosphate-buffered saline, a mass fraction of 1.2?wt% of CaCl₂ flowing in the shell nozzle with a fixed flow rate of 0.08?mL min?1, and a translation velocity of the printing nozzle of 10?mm s?1. To demonstrate the applicability of the technique, preosteoblasts and human adipose stem cells (hASCs) were used to obtain cell-laden structures with multi-layer porous mesh structures. The fabricated cell-laden mesh structures exhibited reasonable initial cell viabilities for preosteoblasts (93%) and hASCs (92%), and hepatogenic differentiation of hASC was successfully achieved.