The effect of recombinant E-cadherin substratum on the differentiation of endoderm-derived hepatocyte-like cells from embryonic stem cells
The effect of recombinant E-cadherin substratum on the differentiation of endoderm-derived hepatocyte-like cells from embryonic stem cells
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DOI：
10.1016/j.biomaterials.2010.11.045
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发表时间：
2011-03-01
期刊：
影响因子：
14
通讯作者：
Akaike, Toshihiro
中科院分区：
文献类型：
--
作者：
Haque, Amranul;Hexig, Bayar;Akaike, Toshihiro
从胚胎干细胞产生特定的细胞谱系是将这些细胞用于临床前应用的先决条件。在这里，我们开发了一种重组E-钙粘素底物，用于在单细胞水平上生成肝祖细胞群。这种人工脱细胞饲养层支持ES细胞逐步分化为具有明确内胚层特征的细胞、肝祖细胞，最后是具有肝细胞表型和功能特征的细胞。肝内胚层细胞的有效分化(约55%)，加上缺乏神经外胚层和中胚层标记，表明选择性地诱导内胚层分化。E-钙粘附素和甲胎蛋白的共同表达(约98%)提示E-钙粘附素作为富集肝前体细胞的表面标志物具有重要作用。通过大量的扩增，可以获得大约92%的白蛋白表达细胞，而不需要任何酶压力和细胞分选。此外，这些小鼠ES细胞来源的肝细胞样细胞在形态上与原代肝细胞有更高的相似性。综上所述，我们证明了E-钙粘素底物可以诱导ES细胞分化为内胚层来源的肝细胞样细胞。该重组细胞外基质即使在单细胞水平上也可以有效地作为研究肝脏早期发育机制的体外模型。(C)2010爱思唯尔有限公司。保留所有权利。
Generation of specific lineages of cells from embryonic stem (ES) cells is pre-requisite to use these cells in pre-clinical applications. Here, we developed a recombinant E-cadherin substratum for generation of hepatic progenitor populations at single cell level. This artificial acellular feeder layer supports the stepwise differentiation of ES cells to cells with characteristics of definitive endoderm, hepatic progenitor cells, and finally cells with phenotypic and functional characteristics of hepatocytes. The efficient differentiation of hepatic endoderm cells (approximately 55%) together with the absence of neuroectoderm and mesoderm markers suggests the selective induction of endoderm differentiation. The co-expression of E-cahderin and alpha-fetoprotein (approximately 98%) suggests the important role of E-cadherin as a surface marker for the enrichment of hepatic progenitor cells. With extensive expansion, approximately 92% albumin expressing cells can be achieved without any enzymatic stress and cell sorting. Furthermore, these mouse ES cell-derived hepatocyte-like cells showed higher morphological similarities to primary hepatocytes. In conclusion, we demonstrated that E-cadherin substratum can guide differentiation of ES cells into endoderm-derived hepatocyte-like cells. This recombinant extracellular matrix could be effectively used as an in vitro model for studying the mechanisms of early stages of liver development even at single cell level. (C) 2010 Elsevier Ltd. All rights reserved.