3D bioprinting of liver spheroids derived from human induced pluripotent stem cells sustain liver function and viability in vitro
3D bioprinting of liver spheroids derived from human induced pluripotent stem cells sustain liver function and viability in vitro
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DOI：
10.1088/1758-5090/ab4a30
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发表时间：
2020-01-01
期刊：
影响因子：
9
通讯作者：
Zatz, Mayana
中科院分区：
文献类型：
--
作者：
Goulart, Ernesto;de Caires-Junior, Luiz Carlos;Zatz, Mayana
肝脏负责许多代谢、内分泌和外分泌功能。每年约有200万例死亡与肝功能衰竭有关。现代3D生物打印技术与自体诱导多能干细胞（iPS）衍生的移植物相结合，可以代表治疗终末期肝病患者的相关组织工程方法。然而，准确概括肝脏的方案？通过生物打印的上皮实质仍然不发达。在这里，我们评估了使用iPS衍生的实质细胞（即肝细胞样细胞）的单细胞分散（即从传统的二维分化获得）与使用iPS衍生的肝细胞样细胞球状体（即三维细胞培养）的影响，两者都与非实质细胞（例如间充质和内皮细胞）组合，进入最终的肝脏组织功能。在培养18天后，与球状体打印的构建体相比，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。此外，单细胞打印的构建体显示上皮-间充质转化，导致肝细胞表型的快速丧失。这些结果表明了使用基于球体的生物打印的优势，有助于改善当前的肝脏生物打印技术，以实现未来的再生医学应用以及肝脏生理学和疾病建模。
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.