Hepatic Differentiation of Liver-Derived Progenitor Cells and Their Characterization by MicroRNA Analysis
Hepatic Differentiation of Liver-Derived Progenitor Cells and Their Characterization by MicroRNA Analysis
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DOI：
10.1002/lt.22111
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发表时间：
2010-09-01
影响因子：
4.6
通讯作者：
Sahin, M. Behnan
中科院分区：
文献类型：
--
作者：
Chen, Yixin;Zhou, Hongchao;Sahin, M. Behnan
我们最近报道了一种称为肝源性祖细胞（LDPCs）的新型祖细胞群的分离和表征，它可以在体外分化为功能性肝细胞。然而，我们最初的研究结果表明，LDPCs的肝分化效率相对较低且多变，没有验证其体内潜能。在这里，我们报道了LDPCs在明确的培养条件下高效和强大的肝分化，以及LDPCs向成熟肝细胞的体内分化。除了形态学研究外，我们还对LDPCs的体外肝分化进行了逆转录聚合酶链反应（RT-PCR）和microRNA分析，以证实分化过程的效率。对分化的LDPCs进行组织学研究发现，50%以上的细胞白蛋白、细胞角蛋白18、肝细胞核因子1 α阳性，并含有糖原颗粒，均符合向功能性肝细胞的分化。我们还通过RT-PCR证明，在分化后，它们表达了成熟肝细胞中发现的几种标记物，并且LDPCs的microRNA谱与新鲜肝细胞的谱相似，证实了我们的形态学发现。最后，将LDPCs移植到二肽基肽酶iv缺陷（DPPIV-/-）大鼠模型中，结果表明LDPCs能够在DPPIV-/-大鼠肝脏中移植并形成成熟的肝细胞。总之，LDPCs是一种独特的肝祖细胞群，能够在体外和体内进行肝分化，这使它们成为重要应用的潜在宝贵资源，如各种肝脏疾病的药理研究和细胞治疗。中华肝病杂志，2010(4):1086-1097。(c) 2010年。
We recently reported the isolation and characterization of a novel population of progenitor cells called liver-derived progenitor cells (LDPCs), which could differentiate into functional hepatocytes in vitro. However, our original studies resulted in relatively low and variable hepatic differentiation efficiency without validation of in vivo potential of LDPCs. Here, we report an efficient and robust hepatic differentiation of LDPCs under well-defined culture conditions and in vivo differentiation of LDPCs to mature hepatocytes. In addition to morphological studies, we performed reverse-transcription polymerase chain reaction (RT-PCR) and microRNA analyses of the in vitro hepatic differentiation of LDPCs to substantiate the efficiency of the differentiation process. The histological studies on the differentiated LDPCs showed that more than 50% of the cells were positive for albumin, cytokeratin 18, and hepatocyte nuclear factor 1 alpha and contained glycogen particles, all consistent with differentiation to functional hepatocytes. We also demonstrated by RT-PCR that upon differentiation, they expressed several markers found in mature hepatocytes and the microRNA profile of LDPCs became similar to the profile of fresh hepatocytes, confirming our morphological findings. Finally, the transplantation of LDPCs in a dipeptidyl peptidase IV-deficient (DPPIV-/-) rat model showed that LDPCs were able to engraft and form mature hepatocytes in the livers of the DPPIV-/- rats. In summary, LDPCs are a unique population of liver progenitor cells capable of hepatic differentiation both in vitro and in vivo, which makes them a potentially valuable resource for important applications such as pharmacological studies and cell therapies for a variety of liver disorders. Liver Transpl 16:1086-1097, 2010. (C) 2010 AASLD.