Treatment of surgically induced acute liver failure by transplantation of HNF4-overexpressing embryonic stem cells.
Treatment of surgically induced acute liver failure by transplantation of HNF4-overexpressing embryonic stem cells.
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DOI：
10.1111/j.1443-9573.2006.00253.x
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发表时间：
2006-01-01
期刊：
影响因子：
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通讯作者：
Xiao, Shu Dong
中科院分区：
文献类型：
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作者：
Kuai, Xiao Ling;Cong, Xiao Qian;Xiao, Shu Dong
目的：分化胚胎干细胞（ES）的组织特异性干细胞具有多能性和遗传灵活性。最近的观察表明，胚胎干细胞可以分化为肝细胞。因此，基于细胞的治疗有可能成为肝移植的一种治疗选择。本研究探讨了过表达肝细胞核因子4 （HNF4）的ES细胞移植治疗大鼠急性肝衰竭的疗效。方法：将HNF4转染到胚胎干细胞中，选择过表达HNF4的胚胎干细胞克隆。逆转录聚合酶链反应（RT-PCR）检测自发分化过表达hnf4的ES细胞中肝细胞分化标志物白蛋白、转甲状腺素、葡萄糖-6-磷酸（G-6-P）和SAPK/ERK激酶-1 (SEK1) mRNA水平。电镜观察自发分化过表达hnf4的胚胎干细胞的超微结构。为了诱导急性肝衰竭，Sprague-Dawley大鼠进行90%肝切除并给予5%葡萄糖口服。老鼠被分成三组。治疗组（n = 12）肝内注射2 × 10(7)个来自同一克隆的未分化过表达hnf4的胚胎干细胞，对照组1 （n = 12）肝内注射2 × 10(7)个未分化胚胎干细胞，对照组2 （n = 12）肝内注射等量培养基，不注射任何细胞。结果：对照组1和对照组2的大鼠均在72 h内死亡，而接受未分化的过表达hnf4的ES细胞移植的大鼠有33%存活超过1个月。自发分化的过表达hnf4的ES细胞仅表达转甲状腺素mRNA。细胞超微结构上富含线粒体和含过氧化氢酶的过氧化物酶体。结论：胚胎干细胞移植可能是急性肝功能不全患者维持生命的潜在治疗方法，并可能成为原位肝移植的桥梁。
OBJECTIVE: Tissue-specific stem cells from differentiating embryonic stem (ES) cells are both pluripotent and genetically flexible. Recent observations indicate that ES cells can differentiate into hepatocytes. Therefore, cell-based therapy can potentially be a therapeutic alternative to liver transplantation. In this study the treatment of acute liver failure in rats by transplantation of hepatocyte nuclear factor 4 (HNF4)-overexpressing ES cells was investigated.METHODS: The HNF4 was transfected into ES cells and ES cell clones overexpressing HNF4 were selected. The levels of markers of hepatocyte differentiation, including albumin, transthyretin, glucose-6-phosphates (G-6-P) and SAPK/ERK kinase-1 (SEK1) mRNA, were tested in spontaneously differentiated HNF4-overexpressing ES cells by reverse transcription-polymerase chain reaction (RT-PCR). The ultrastructure of the spontaneously differentiated HNF4-overexpressing ES cells was examined by electron microscopy. To induce acute liver failure, Sprague-Dawley rats were subjected to 90% hepatectomy and given 5% oral dextrose. The rats were divided into three groups. The rats in the treatment group (n = 12) received intraliver injection of 2 x 10(7) undifferentiated HNF4-overexpressing ES cells from the same clone, the rats in control group 1 (n = 12) received 2 x 10(7) undifferentiated ES cells, and the rats in control group 2 (n = 12) received the same volume of media without any cells.RESULTS: All rats in control group 1 and control group 2 died within 72 h, while 33% of rats that received undifferentiated HNF4-overexpressing ES cells transplantation survived more than 1 month. Spontaneously differentiated HNF4-overexpressing ES cells only expressed transthyretin mRNA. The cells were rich in mitochondrion and catalase-containing peroxisomes in ultrastructure.CONCLUSIONS: Transplantation of ES cells could be a potential treatment in supporting life during acute liver insufficiency and could be a bridge to orthotopic liver transplantation.