Generation of hepatocytes from cultured mouse embryonic stem cells
Generation of hepatocytes from cultured mouse embryonic stem cells
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DOI：
10.1053/jlts.2003.50207
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发表时间：
2003-10-01
影响因子：
4.6
通讯作者：
Xiao, SD
中科院分区：
文献类型：
--
作者：
Kuai, XL;Cong, XQ;Xiao, SD
胚胎干细胞（ES）是从体外受精的囊胚内细胞群中获得的多能细胞。胚胎干细胞可以被诱导分化成潜在的所有细胞类型。本研究的目的是检测小鼠胚胎干细胞在维甲酸（RA）、肝细胞生长因子（HGF）和-神经生长因子（β - ngf）存在下向肝细胞分化的潜力。在细胞培养中加入RA、HGF和- ngf。通过α(1)-抗胰蛋白酶（α (1)-AT）和α(1)-蛋白（AFP）表达的免疫组化检测和白蛋白、转甲状腺素、葡萄糖6磷酸、肝核因子4和SAPK/ERK激酶1 （SEK1）信使RNA的逆转录酶聚合酶链反应试验，从形态学和生化角度证实了肝细胞的诱导作用。在HGF和β - ngf加入细胞培养15天后，可以观察到许多上皮样细胞。同时检测到α (1)-AT、AFP、白蛋白、转甲状腺素、葡萄糖6磷酸、肝核因子4和SEK1信使RNA的表达，表明ES细胞成功分化为功能肝细胞。然而，在单独存在RA时，只有转甲状腺素信使RNA阳性，而未检测到与功能肝细胞相关的其他表达。在HGF和β - ngf存在的情况下，小鼠胚胎干细胞可以分化为功能性肝细胞，而RA功能则受到限制。
Embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of fertilized blastocysts in vitro. ES cells can be induced to undergo differentiation into potentially all cell types. The aim of this study is to examine the differentiating potential of mouse ES cells into hepatocytes in the presence of retinoic acid (RA), hepatocyte growth factor (HGF), and beta-nerve growth factor (beta-NGF). RA, HGF, and beta-NGF were added to the cell culture. Hepatocyte induction was confirmed morphologically, as well as biochemically, through immunohistochemical assays of alpha(1)-antitrypsin (alpha(1)-AT) and alfafeta-protein (AFP) expression and reverse-transcriptase polymerase chain reaction tests for the presence of albumin, transthyretin, glucose 6 phosphates, hepatic nuclear factor 4, and SAPK/ERK kinase-1 (SEK1) messenger RNA, produced only by functioning hepatocytes. Fifteen days after the addition of HGF and beta-NGF to the cell culture, many epithelioid cells were noticed. alpha(1)-AT, AFP, albumin, transthyretin, glucose 6 phosphates, hepatic nuclear factor 4, and SEK1 messenger RNA expression also was detected, indicating successful ES cell differentiation into functioning hepatocytes. However, in the presence of RA alone, only transthyretin messenger RNA was positive, whereas no other expression pertaining to functioning hepatocytes could be detected. In the presence of HGF and beta-NGF, mouse ES cells can differentiate into functioning hepatocytes, whereas RA function is limited.