Defined conditions for development of functional hepatic cells from human embryonic stem cells
Defined conditions for development of functional hepatic cells from human embryonic stem cells
复制标题
DOI：
10.1089/scd.2005.14.643
复制
发表时间：
2005-12-01
影响因子：
4
通讯作者：
Kaufman, DS
中科院分区：
文献类型：
--
作者：
Schwartz, RE;Linehan, JL;Kaufman, DS
人类胚胎干细胞（hES）为评估调节特定细胞系发育的特异性可溶性和细胞结合刺激提供了重要手段。在这里，我们研究了支持hES细胞向肝细胞分化的特定细胞因子和细胞外基质（ECM）蛋白。几种不同条件下的试验表明，在完全无血清培养的he细胞衍生的胚状体中添加成纤维细胞生长因子(FGF)-4和肝细胞生长因子，随后允许其附着在I型胶原包被的培养皿上，从而最大限度地分化为细胞，不仅具有肝细胞的形态和表型特征，而且具有功能特征。在这些条件下，常见的肝脏转录因子HNF-3 β、HNF-1和GATA-4的表达均被显著诱导。肝细胞功能通过多种互补标准得到证实：尿素和白蛋白的产生、苯巴比妥诱导的细胞色素P450的表达和吲哚菁绿的摄取。这些hES细胞衍生的肝细胞将作为了解正常人类肝细胞发育的资源，并用于细胞替代疗法和药物筛选等应用。
Human embryonic stem (hES) cells provide an important means to evaluate specific soluble and cell-bound stimuli that regulate development of specific cell lineages. Here, we examined specific cytokines and extracellular matrix (ECM) proteins that support differentiation of hES cells to hepatocytes. Tests of several different conditions determined that addition of fibroblast growth factor (FGF)-4 and hepatocyte growth factor in completely serum-free cultures of hES cell-derived embryoid bodies subsequently allowed to attach to type I collagen-coated dishes led to maximal differentiation into cells, not only with the morphologic and phenotypic characteristics of hepatocytes but also the functional characteristics. Expression of common hepatic transcription factors including HNF-3 beta, HNF-1, and GATA-4 were all significantly induced under these conditions. Hepatocyte function was demonstrated by multiple complementary criteria: production of urea and albumin, phenobarbital-induced cytochrome P450 expression, and uptake of indocyanine green. These hES cell-derived hepatocytes will serve as a resource to understand normal human hepatocyte development and for applications such as cell replacement therapies and screening of pharmacologic drugs.