Wang L-Y, Liu L-P, Ge J-Y, Yuan Y-Y, Sun L-L, Xu H, Huang P-Y, Hui L-J, Isoda H, Ohkohchi N, Li Y-M, Zheng Y-W
Research Center of Stem Cell and Regenerative Medicine, Affiliated Hospital of Jiangsu University, Zhenjiang, China; Department of Dermatology, Affiliated Hospital of Jiangsu University, Zhenjiang, China.
Faculty of Medicine, University of Tsukuba, Tsukuba, Japan.
Transplant Proc. 2018 Nov;50(9):2864-2867. doi: 10.1016/j.transproceed.2018.03.076. Epub 2018 Mar 15.
The difficulty in proliferation and availability and the rapid loss functions of primary human hepatocytes highlight the need to develop an alternative, preferably renewable source of human induced hepatocytes in regenerative medicine. Liver organoids generated on a multiple-cell microenvironment in a 3-dimensional (3D) system can provide a highly efficient solution to this issue.
Human hepatocytes were induced from fibroblasts by the lentiviral expression of FOXA3, HNF1A, and HNF4A. Together with these induced hepatocytes, human umbilical vein endothelial cells and mesenchymal stem cells in a 3D system were used to produce liver organoids. Liver-related gene and protein expression of liver organoids and induced hepatocytes were tested using a 2-dimensional (2D) system.
Liver organoids notably increased the expression of hepatic transcription factors, marker genes, transporter genes, and liver metabolism enzyme genes, while it decreased the specific gene expression of fibroblasts. Liver organoids expressed comparable liver-specific proteins, such as ALB, AAT, and HNF4A in the 3D system.
Direct reprogramming in multiple-cell microenvironments in 3D systems is more controllable and efficient than cell reprogramming in 2D systems. Liver organoids have the potential for use in disease modeling, pharmaceutical applications, and cellular transplantation.
原代人肝细胞增殖困难、获取不易且功能快速丧失，这凸显了在再生医学中开发替代的、最好是可再生的人诱导肝细胞来源的必要性。在三维（3D）系统的多细胞微环境中生成的肝脏类器官可为该问题提供高效解决方案。
通过慢病毒表达FOXA3、HNF1A和HNF4A，将成纤维细胞诱导为人肝细胞。将这些诱导的肝细胞与人脐静脉内皮细胞和间充质干细胞一起用于在3D系统中生成肝脏类器官。使用二维（2D）系统检测肝脏类器官和诱导肝细胞的肝脏相关基因及蛋白质表达。
肝脏类器官显著增加了肝转录因子、标记基因、转运蛋白基因和肝脏代谢酶基因的表达，同时降低了成纤维细胞的特异性基因表达。肝脏类器官在3D系统中表达了相当水平的肝脏特异性蛋白质，如ALB、AAT和HNF4A。
在3D系统的多细胞微环境中进行直接重编程比在2D系统中进行细胞重编程更可控、更高效。肝脏类器官具有用于疾病建模、药物应用和细胞移植的潜力。
