Department of Chemical Engineering, Faculty of Engineering, Kyushu University, Fukuoka, Japan.
Tissue Eng Part A. 2011 Aug;17(15-16):2071-8. doi: 10.1089/ten.TEA.2010.0689. Epub 2011 May 11.
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.
我们专注于多能干细胞作为混合型人工肝（HAL）的潜在来源，并试图开发一种将多能干细胞分化为肝谱系细胞的方法。在这项研究中，我们通过应用中空纤维（HF）/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）的肝分化，其中培养的细胞在 HF 的管腔中形成称为“类器官”的细胞聚集体。将 ES 和 iPS 细胞注入 HF 中以诱导类器官形成，并进行细胞培养。为了诱导肝分化，我们向培养基中添加了促进分化的试剂。分化相关基因的表达水平上调，HF 内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，因此分化的 ES 和 iPS 细胞的每单位体积的肝特异性功能的最高水平达到了与原代小鼠肝细胞相当或更好的水平。总之，ES 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开发 HAL 的基础技术。
