Hepatocyte-like cells derived from human embryonic stem cells specifically via definitive endoderm and a progenitor stage
Hepatocyte-like cells derived from human embryonic stem cells specifically via definitive endoderm and a progenitor stage
复制标题
DOI：
10.1016/j.jbiotec.2009.11.007
复制
发表时间：
2010-02-01
影响因子：
4.1
通讯作者：
Heins, Nico
中科院分区：
文献类型：
--
作者：
Brolen, Gabriella;Sivertsson, Louise;Heins, Nico
人胚胎干细胞为功能性肝细胞提供了潜在的无限供应，因为它们可以分化成肝细胞样细胞，显示出特征性的肝脏形态并表达各种肝脏标志物。这些细胞可用于各种应用，例如药物代谢和肝毒性的研究，然而，这将需要药物代谢酶的显着表达。为了获得这些细胞，我们使用逐步分化方案，其中添加生长和成熟因子。第一阶段涉及定形内胚层的形成。接下来，用已知促进肝祖细胞类型的诱导和增殖的因子处理这些细胞。在最后阶段，细胞终末分化并成熟为功能性肝细胞样细胞。通过分析内胚层或肝标志物表征培养物，并与未经定形内胚层诱导的培养物进行比较。肝脏功能，如尿素分泌，糖原储存，吲哚菁绿色的吸收和分泌，细胞色素P450的表达和活性进行了评价。DE-Hep显示出亚组织细胞的肝细胞形态，并表现出许多肝脏功能，包括转运活性和代谢重要的细胞色素P450亚家族特异性药物的能力。这是hESC向功能性肝细胞分化的重要步骤。(C)2009爱思唯尔有限公司版权所有。
Human embryonic stem cells offer a potential unlimited supply for functional hepatocytes, since they can differentiate into hepatocyte-like cells displaying a characteristic hepatic morphology and expressing various hepatic markers. These cells could be used in various applications such as studies of drug metabolism and hepatotoxicity, which however, would require a significant expression of drug metabolizing enzymes. To derive these cells we use a stepwise differentiation protocol where growth- and maturation factors are added. The first phase involves the formation of definitive endoderm. Next, these cells are treated with factors known to promote the induction and proliferation towards hepatic progenitor cell types. In the last phase the cells are terminally differentiated and maturated into functional hepatocyte-like cells. The cultures were characterized by analysis of endodermal or hepatic markers and compared to cultures derived without induction via definitive endoderm. Hepatic functions such as urea secretion, glycogen storage, indocyanine green uptake and secretion, and cytochrome P450-expression and activity were evaluated.The DE-Hep showed a hepatocyte morphology with sub-organized cells and exhibited many liver-functions including transporter activity and capacity to metabolize drugs specific for important cytochrome P450 sub-families. This represents an importantstep in differentiation of hESC into functional hepatocytes. (C) 2009 Elsevier B.V. All rights reserved.