In vitro differentiation of hepatocyte-like cells from embryonic stem cells promoted by gene transfer of hepatocyte nuclear factor 3 β

被引:32
|
作者
Kanda, S
Shiroi, A
Ouji, Y
Birumachi, J
Ueda, S
Fukui, H
Tatsumi, K
Ishizaka, S
Takahashi, Y
Yoshikawa, M
机构
[1] Nara Med Univ, Div Dev Biol, Dept Parasitol, Kashihara, Nara 6348521, Japan
[2] Nara Med Univ, Dept Internal Med 3, Kashihara, Nara 6348521, Japan
[3] Nara Med Univ, Dept Anat 2, Kashihara, Nara 6348521, Japan
[4] Nara Inst Sci & Technol, Grad Sch Biol Sci, Nara 6300101, Japan
关键词
ES cells; HNF-3; beta; embryoid bodies; hepatic differentiation;
D O I
10.1016/S1386-6346(03)00114-1
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
We investigated the ability of a genetically altered embryonic stem (ES) cell line to promote endodermal differentiation toward hepatocytes in vitro by transfecting the hepatocyte nuclear factor-3beta (HNF-3beta) gene. Parental and HNF-3beta-transfected ES cells were initiated toward differentiation in embryoid bodies (EBs) for 5 days and the resulting EBs were transferred to an attached culture system. Albumin production was observed using an immuno-cytochemical method 7 days after induction of differentiation in almost all differentiating HNF-3beta-transfected ES cells, whereas scant immuno-reactivity against albumin was found on the same day in the cultures of differentiating parental ES cells. An analysis using a reverse transcriptase polymerase chain reaction revealed the HNF-4alpha expression in the HNF-3beta-transfected ES cells and also demonstrated that the expression of endodermal and hepatocyte-related markers, such as transthyretin, alpha-fetoprotein, albumin, alpha-1 antitrypsin, tryptophan-2,3-dioxygenase and phosphoenol-pyruvate carboxykinase, could be observed at an early stage in the outgrowths of HNF-3beta-transfected ES cells compared to the parental ES cells. These results suggest that HNF-3beta-transfected ES cells may be useful for the efficient induction of hepatocytes in vivo. (C) 2003 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:225 / 231
页数:7
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