Wellcome Trust–Medical Research Council Stem Cell Institute, Anne McLaren Laboratory for Regenerative Medicine, Department of Surgery, University of Cambridge, Cambridge, UK.
Nat Protoc. 2013 Feb;8(2):430-7. doi: 10.1038/nprot.2012.153.
Large-scale production of hepatocytes from a variety of genetic backgrounds would be beneficial for drug screening and to provide a source of cells to be used as a substitute for liver transplantation. However, fully functional primary hepatocytes remain difficult to expand in vitro, and circumventing this problem by using an alternative source of cells is desirable. Here we describe a 25-d protocol to direct the differentiation of human pluripotent stem cells into a near-homogenous population of hepatocyte-like cells. As cells progress through this protocol, they express genes in a chronological manner similar to that described during in vivo hepatic development. The protocol relies on culture systems devoid of serum, feeders or complex extracellular matrices, which enable molecular analyses without interference from unknown factors. This approach works efficiently with human embryonic stem cells and human induced pluripotent stem cells and was recently used to model liver diseases in vitro.
从各种遗传背景大量生产肝细胞将有利于药物筛选，并提供可替代肝移植的细胞来源。然而，完全功能的原代肝细胞在体外仍难以扩增，因此理想的方法是利用替代细胞来源来解决这个问题。在这里，我们描述了一个 25 天的方案，可将人多能干细胞定向分化为近同质的肝细胞样细胞群体。随着细胞通过这个方案的进展，它们以类似于体内肝发育过程中描述的时间顺序表达基因。该方案依赖于无血清、饲养层或复杂细胞外基质的培养系统，这使得能够在不受未知因素干扰的情况下进行分子分析。该方法与人类胚胎干细胞和人类诱导多能干细胞高效配合，并最近被用于体外模拟肝脏疾病。
