Hay David C, Zhao Debiao, Ross Arlene, Mandalam Ramkumar, Lebkowski Jane, Cui Wei
Department of Gene Function and Development, Roslin Institute, Roslin, Midlothian, Scotland, United Kingdom.
Cloning Stem Cells. 2007 Spring;9(1):51-62. doi: 10.1089/clo.2006.0045.
The utilization of human hepatocytes for biomedical research, drug discovery, and treatment of liver diseases is hindered by the limited availability of donated livers and the variability of their derived hepatocytes. Human embryonic stem cells (hESCs) are pluripotent and provide a unique, unlimited resource for human hepatocytes. However, differentiation of hESCs to hepatocytes remains a challenge. We have developed a multistage procedure by which hESCs can be directly differentiated to hepatocyte-like cells without embryoid body formation and the requirement of sodium butyrate. The hESC-derived hepatocyte-like cells (HLCs) exhibited characteristic hepatocyte morphology, expressed hepatocyte markers, including alpha-fetoprotein, albumin, and hepatocyte nuclear factor 4alpha, and possessed hepatocyte-specific activities, such as p450 metabolism, albumin production, glycogen storage, and uptake and excretion of indocyanine green. Hepatocyte growth factor was found to play a positive role in promoting hepatocyte differentiation. Our differentiation system has shown that hESCs can be differentiated to hepatocyte-like cells capable of executing a range of hepatocyte functions. Therefore, it presents a proof-of-principle of potential applications of using the hESC-derived hepatocytes. Additionally, the hESC-derived HLCs provide a unique model to study the mechanisms involved in human hepatocyte differentiation and liver function.
用于生物医学研究、药物研发以及肝脏疾病治疗的人肝细胞，因捐赠肝脏的有限可用性及其所衍生肝细胞的变异性而受到阻碍。人胚胎干细胞（hESCs）具有多能性，为人类肝细胞提供了独特、无限的资源。然而，将hESCs分化为肝细胞仍然是一项挑战。我们开发了一种多阶段程序，通过该程序，hESCs可以直接分化为类肝细胞，而无需形成胚状体且无需丁酸钠。hESC衍生的类肝细胞（HLCs）呈现出特征性的肝细胞形态，表达肝细胞标志物，包括甲胎蛋白、白蛋白和肝细胞核因子4α，并具有肝细胞特异性活性，如p450代谢、白蛋白产生、糖原储存以及吲哚菁绿的摄取和排泄。发现肝细胞生长因子在促进肝细胞分化中起积极作用。我们的分化系统表明，hESCs可以分化为能够执行一系列肝细胞功能的类肝细胞。因此，它为使用hESC衍生的肝细胞的潜在应用提供了原理证明。此外，hESC衍生的HLCs提供了一个独特的模型来研究人类肝细胞分化和肝功能所涉及的机制。
