Murine embryonic stem cell-derived hepatic progenitor cells engraft in recipient livers with limited capacity of liver tissue formation
Murine embryonic stem cell-derived hepatic progenitor cells engraft in recipient livers with limited capacity of liver tissue formation
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DOI：
10.3727/096368908784153896
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发表时间：
2008-01-01
影响因子：
3.3
通讯作者：
Ott, Michael
中科院分区：
文献类型：
--
作者：
Sharma, Amar Deep;Cantz, Tobias;Ott, Michael
小鼠胚胎干细胞（ES）的定向内胚层分化产生具有肝表型的细胞亚群。这种ES细胞来源的肝祖细胞（ES-HPC）可以在体外获得肝细胞的特征，但不能在体内形成实质性的肝细胞簇。在这项研究中，我们调查了这是否是由于植入效率低下或不成熟的表型ES-HPC。ES细胞移植到NOD/SCID小鼠的受体肝脏中，28天后具有与成年肝细胞相似的功效。由于移植的未纯化的ES-HPC在脾和肝中形成畸胎瘤，我们应用白蛋白启动子/增强子驱动的报告系统通过细胞分选纯化ES-HPC。肝细胞特异性基因的RT-PCR分析表明，这些细胞表现出肝脏表型，缺乏多能性标记Oct 4的表达，与11.5天胚胎的细胞相当。将来自β-半乳糖苷酶转基因ES细胞的分选的ES-HPC注射到富马酰乙酰乙酸缺陷（FAH（-/-））SCID小鼠中，并在8至12周后进行分析。用X-gal溶液染色显示整个肝脏中存在移植细胞。然而，FAH蛋白的免疫组化染色表明肝细胞形成的频率非常低，没有大的肝细胞簇形成的证据。总之，有限的再增殖能力的ES-HPC是不是由一个失败的主要植入，但可能是由于一个不成熟的肝表型移植的ES-HPC。
Directed endodermal differentiation of murine embryonic stem (ES) cells gives rise to a subset of cells with a hepatic phenotype. Such ES cell-derived hepatic progenitor cells (ES-HPC) can acquire features of hepatocytes in vitro, but fail to form substantial hepatocyte clusters in vivo. In this study, we investigated whether this is due to inefficient engraftment or an immature phenotype of ES-HPC. ES cells engrafted into recipient livers of NOD/SCID mice with a similar efficacy as adult hepatocytes after 28 days. Because transplanted unpurified ES-HPC formed teratomas in the spleen and liver, we applied an albumin promoter/enhancer-driven reporter system to purify ES-HPC by cell sorting. RT-PCR analyses for hepatocyte-specific genes showed that the cells exhibited a hepatic phenotype, lacking the expression of the pluripotency marker Oct4, comparable to cells of day 11.5 embryos. Sorted ES-HPC derived from P-galactosidase transgenic ES cells were injected into fumaryl-acetoacetate-deficient (FAH(-/-)) SCID mice and analyzed after 8 to 12 weeks. Staining with X-gal solution revealed the presence of engrafted cells throughout the liver. However, inununostaining for the FAH protein indicated hepatocyte formation at a very low frequency, without evidence for large hepatocyte cluster formation. In conclusion, the limited repopulation capacity of ES-HPC is not caused by a failure of primary engraftment, but may be due to an immature hepatic phenotype of the transplanted ES-HPC.