Hepatic Differentiation of Mouse Embryonic Stem Cells and Induced Pluripotent Stem Cells During Organoid Formation in Hollow Fibers
Hepatic Differentiation of Mouse Embryonic Stem Cells and Induced Pluripotent Stem Cells During Organoid Formation in Hollow Fibers
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DOI：
10.1089/ten.tea.2010.0689
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发表时间：
2011-08-01
影响因子：
4.1
通讯作者：
Kajiwara, Toshihisa
中科院分区：
文献类型：
--
作者：
Amimoto, Naoki;Mizumoto, Hiroshi;Kajiwara, Toshihisa
我们关注多能干细胞作为杂交型人工肝脏（HAL）的潜在来源，并试图开发一种将多能干细胞分化为肝细胞谱系的方法。在本研究中，我们采用中空纤维/类器官培养方法，研究了小鼠胚胎干细胞（ES）和诱导多能干细胞（iPS）的肝脏分化，培养的细胞在HF管腔中形成称为“类器官”的细胞聚集体。将胚胎干细胞和诱导性多能干细胞注入HFs诱导类器官形成，并进行细胞培养。为了诱导肝细胞分化，我们在培养基中加入促分化剂。分化相关基因表达水平上调，细胞增殖和类器官形成。由于我们能够在培养中实现高细胞密度，分化的ES和iPS细胞每单位体积的最大肝脏特异性功能水平达到与小鼠原代肝细胞相当或更好的水平。综上所述，胚胎干细胞和iPS细胞有可能成为HAL的细胞来源，因此HF/类器官培养方法有望成为HAL开发的基础技术。
We have focused on pluripotent stem cells as a potential source of a hybrid-type artificial liver (HAL) and tried to develop a method for differentiating the pluripotent stem cells into cells of a hepatic lineage. In this study, we investigated the hepatic differentiation of mouse embryonic stem (ES) cells and induced pluripotent stem (iPS) cells by applying hollow fiber (HF)/organoid culture method, in which cultured cells form a cellular aggregate called an "organoid" in the lumen of the HF. ES and iPS cells were injected into HFs to induce organoid formation, and cells were cultured. To induce hepatic differentiation, we added differentiation-promoting agents to the culture medium. The expression levels of differentiation-related genes were up-regulated, with cell proliferation and organoid formation inside HFs. Since we were able to achieve a high cell density in culture, the maximum levels of liver-specific functions per unit volume in the differentiating ES and iPS cells reached a level comparable to or better than that of primary mouse hepatocytes. In conclusion, ES and iPS cells have the potential to be a cell source for a HAL, and the HF/organoid culture method, therefore, has promise as a basic technology for the development of a HAL.