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

一种新型细胞打印方法及其在人脂肪干细胞嵌入网状结构肝细胞分化中的应用

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作者:Seung Hyun Ahn ,Hyeong Jin Lee ,Ji-Seon Lee ,Hyeon Yoon ,Wook Chun ,Geun Hyung Kim

Abstract

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 × 10(7) 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.

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