Isolation and transcriptional profiling of purified hepatic cells derived from human embryonic stem cells
Isolation and transcriptional profiling of purified hepatic cells derived from human embryonic stem cells
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DOI：
10.1634/stemcells.2007-0964
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发表时间：
2008-08-01
期刊：
影响因子：
5.2
通讯作者：
Baker, Julie
中科院分区：
文献类型：
--
作者：
Chiao, Eric;Elazar, Menashe;Baker, Julie
人胚胎干细胞(HESCs)分化为功能性肝细胞为研究肝发育的分子机制提供了强大的体外模型系统。此外，从hESCs分化而来的具有良好特性的可再生肝细胞可用于体外药物代谢和毒理学分析，筛选潜在的抗病毒药物，以及基于细胞的治疗肝病的方法。在这项研究中，我们描述了一种将hESCs分化为具有复杂细胞形态的肝细胞的方案。用慢病毒载体鉴定和分离可能的肝细胞，该慢病毒载体含有驱动增强型绿色荧光蛋白表达的甲胎蛋白启动子(AFP：EGFP)。使用最近发布的Affymetrix外显子阵列ST1.0(Santa Clara，CA，http://www.affymetrix.com).)对AFP：EGFP+和AFP：EGFP-细胞群体的三个样本进行全基因组转录图谱对转录图谱的统计分析表明，AFP：EGFP+群体的肝细胞特征基因高度丰富。这些数据提供了对复杂的肝细胞分化过程的独特见解，指出了可以更有效地指导hESCs向成熟肝细胞分化的信号通路，并确定了可能用于进一步分离hESCs向肝细胞分化的分子标记。
The differentiation of human embryonic stem cells (hESCs) into functional hepatocytes provides a powerful in vitro model system for studying the molecular mechanisms governing liver development. Furthermore, a well-characterized renewable supply of hepatocytes differentiated from hESCs could be used for in vitro assays of drug metabolism and toxicology, screening of potential antiviral agents, and cell-based therapies to treat liver disease. In this study, we describe a protocol for the differentiation of hESCs toward hepatic cells with complex cellular morphologies. Putative hepatic cells were identified and isolated using a lentiviral vector, containing the alpha-feto-protein promoter driving enhanced green fluorescent protein expression (AFP:eGFP). Whole-genome transcriptional profiling was performed on triplicate samples of AFP:eGFP+ and AFP:eGFP- cell populations using the recently released Affymetrix Exon Array ST 1.0 (Santa Clara, CA, http://www.affymetrix.com). Statistical analysis of the transcriptional profiles demonstrated that the AFP:eGFP+ population is highly enriched for genes characteristic of hepatic cells. These data provide a unique insight into the complex process of hepatocyte differentiation, point to signaling pathways that may be manipulated to more efficiently direct the differentiation of hESCs toward mature hepatocytes, and identify molecular markers that may be used for further dissection of hepatic cell differentiation from hESCs.