Hypoxia efficiently induces differentiation of mouse embryonic stem cells into endodermal and hepatic progenitor cells
Hypoxia efficiently induces differentiation of mouse embryonic stem cells into endodermal and hepatic progenitor cells
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DOI：
10.1016/j.bej.2013.02.012
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发表时间：
2013-05-15
影响因子：
3.9
通讯作者：
Sakai, Yasuyuki
中科院分区：
文献类型：
--
作者：
Katsuda, Takeshi;Teratani, Takumi;Sakai, Yasuyuki
虽然ES细胞对基于细胞的肝病治疗具有潜在的影响，但其有效分化为功能性肝细胞仍然困难。解决这个问题的一个可能的方法是控制培养环境以重现体内肝脏发育。考虑到胚胎在其早期阶段暴露于低氧条件，低氧条件似乎适合于诱导ES细胞的早期分化。然而，还没有研究评估缺氧对ES细胞的内胚层或早期肝分化的影响。在这里，我们研究了缺氧作为一种调节因子在小鼠ES细胞的内胚层和随后的早期肝分化的潜在作用。在维甲酸的刺激下，典型的内胚层形态在缺氧（5%O-2）条件下比常氧条件下更明显。因此，定量RT-PCR显示Foxa 2和GATA 4，内胚层特化的必需转录因子的显着增加。此外，缺氧放大肝诱导因子FGF 1、FGF 4和HGF的作用，这通过显著增加的ALB-GFP阳性细胞分数以及增加的ALB、AFP和CK 18的基因表达水平来证明。低氧诱导的肝前体细胞可高效分化为具有分泌ALB能力的成熟肝细胞。我们还发现，分化细胞不消耗氧气，即使在常氧，这表明氧气本身可能有一个正常的内胚层和早期肝分化的ES细胞的负面影响。这些结果将为肝祖细胞的高效生产提供有用的见解，并有助于基于干细胞的肝组织工程。(C)2013爱思唯尔有限公司版权所有。
Although ES cells have potential impact on cell-based therapy for liver diseases, their efficient differentiation into functional hepatocytes remains difficult. One possible approach to this issue is to control the culture environment to recapitulate the in vivo liver development. Given that the embryo is exposed to hypoxic condition in its early stage, hypoxic condition seems suitable for inducing early differentiation of ES cells. However, no studies have evaluated the effect of hypoxia on endodermal or early hepatic differentiation of ES cells. Here we investigated the potential role of hypoxia as a regulatory factor in endodermal and subsequent early hepatic differentiation of mouse ES cells. Under the stimulus of retinoic acid, a typical endodermal morphology emerged more dominantly under hypoxia (5% O-2) than normoxia. Accordingly, quantitative RT-PCR revealed significant increase in Foxa2 and GATA4, essential transcription factors for endoderm specification. In addition, hypoxia amplified the effects of hepatic induction factors FGF1, FGF4 and HGF, which was evidenced by dramatically increased ALB-GFP-positive cell fraction, as well as increased gene expression levels of ALB, AFP, and CK18. Furthermore, the hepatic progenitor cells induced under hypoxic condition were efficiently differentiated into mature hepatocytes with ALB secretion ability. We also found that the differentiating cells did not consume oxygen even under normoxia, suggesting that oxygen per se might have a negative effect on the normal endodermal and early hepatic differentiation of ES cells. These results will provide useful insight into efficient production of hepatic progenitor cells, and contribute to stem cell-based liver tissue engineering. (C) 2013 Elsevier B.V. All rights reserved.