The liver is responsible for many metabolic, endocrine and exocrine functions. Approximately 2 million deaths per year are associated with liver failure. Modern 3D bioprinting technologies allied with autologous induced pluripotent stem cells (iPS)-derived grafts could represent a relevant tissue engineering approach to treat end stage liver disease patients. However, protocols that accurately recapitulates liver's epithelial parenchyma through bioprinting are still underdeveloped. Here we evaluated the impacts of using single cell dispersion (i.e. obtained from conventional bidimensional differentiation) of iPS-derived parenchymal (i.e. hepatocyte-like cells) versus using iPS-derived hepatocyte-like cells spheroids (i.e. three-dimensional cell culture), both in combination with non-parenchymal cells (e.g. mesenchymal and endothelial cells), into final liver tissue functionality. Single cell constructs showed reduced cell survival and hepatic function and unbalanced protein/amino acid metabolism when compared to spheroid printed constructs after 18 days in culture. In addition, single cell printed constructs revealed epithelial-mesenchymal transition, resulting in rapid loss of hepatocyte phenotype. These results indicates the advantage of using spheroid-based bioprinting, contributing to improve current liver bioprinting technology towards future regenerative medicine applications and liver physiology and disease modeling.

中文翻译：

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源自人类诱导的多能干细胞的肝球体的3D生物打印在体外维持肝功能和生存能力。

肝脏负责许多新陈代谢，内分泌和外分泌功能。每年约有200万人死于肝衰竭。与自体诱导多能干细胞（iPS）衍生的移植物结合的现代3D生物打印技术可能代表了一种治疗晚期肝病患者的相关组织工程方法。然而，通过生物打印准确概括肝脏上皮实质的方案仍未开发。在这里，我们评估了使用iPS来源的实质（即肝细胞样细胞）的单细胞分散（即从常规的二维分化获得）与使用iPS来源的类肝细胞球体（即三维细胞培养物）相比的影响。结合非实质细胞（例如间充质和内皮细胞），进入最终的肝组织功能。与培养18天后的球状印刷构建体相比，单细胞构建体显示出降低的细胞存活率和肝功能以及不平衡的蛋白质/氨基酸代谢。另外，单细胞印刷的构建体显示上皮-间充质转化，导致肝细胞表型快速丧失。这些结果表明使用基于球体的生物打印技术的优势，有助于改进当前的肝脏生物打印技术，以用于未来的再生医学应用以及肝脏生理学和疾病建模。单细胞印刷的构建体显示上皮-间质转化，导致肝细胞表型快速丧失。这些结果表明使用基于球体的生物打印技术的优势，有助于改进当前的肝脏生物打印技术，以用于未来的再生医学应用以及肝脏生理学和疾病建模。单细胞印刷的构建体显示上皮-间质转化，导致肝细胞表型快速丧失。这些结果表明使用基于球体的生物打印技术的优势，有助于改进当前的肝脏生物打印技术，以用于未来的再生医学应用以及肝脏生理学和疾病建模。