Evaluation of a Hybrid Artificial Liver Module Based on a Spheroid Culture System of Embryonic Stem Cell-Derived Hepatic Cells
Evaluation of a Hybrid Artificial Liver Module Based on a Spheroid Culture System of Embryonic Stem Cell-Derived Hepatic Cells
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DOI：
10.3727/096368911x605321
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发表时间：
2012-01-01
影响因子：
3.3
通讯作者：
Kajiwara, Toshihisa
中科院分区：
文献类型：
--
作者：
Mizumoto, Hiroshi;Hayashi, Shunsuke;Kajiwara, Toshihisa
混合人工肝（HAL）是一种体外循环系统，由含有固定功能肝细胞的生物反应器组成。预计它不仅可以作为临时的肝功能支持系统，还可以加速肝衰竭恢复过程中的肝脏再生。开发混合型人工肝最困难的问题之一是获得足够的细胞来源。在这项研究中，我们尝试使用聚氨酯泡沫（PUF）/球体培养物通过肝谱系区分胚胎干（ES）细胞，其中培养的细胞在PUF的孔中自发形成球形多细胞聚集体（球体）。我们还通过在体外和离体实验中将 ES 细胞与原代肝细胞进行比较来证明 PUF-HAL 系统的可行性。小鼠 ES 细胞在 PUF 的孔中形成多细胞球体。在用分化促进剂丁酸钠（SB）处理后，ES细胞表达了肝脏特异性功能（氨去除和白蛋白分泌）。我们设计了一个 PUF-HAL 模块，由一个圆柱形 PUF 块和许多中等流量毛细管组成，用于 ES 细胞的肝分化。 SB培养2周后，PUF-HAL模块细胞表现出氨去除和白蛋白分泌功能。由于ES细胞的高增殖活性和高细胞密度，模块单位体积白蛋白分泌功能的最大表达水平与原代小鼠肝细胞培养中所见的相当。在大鼠动物实验中，PUF-HAL 分化的 ES 细胞似乎对肝衰竭的恢复有部分贡献。这一结果表明，含有分化 ES 细胞的 PUF 模块可能是混合人工肝支持系统的有用生物组件。
Hybrid artificial liver (HAL) is an extracorporeal circulation system comprised of a bioreactor containing immobilized functional liver cells. It is expected to not only serve as a temporary liver function support system, but also to accelerate liver regeneration in recovery from hepatic failure. One of the most difficult problems in developing a hybrid artificial liver is obtaining an adequate cell source. In this study, we attempt to differentiate embryonic stem (ES) cells by hepatic lineage using a polyurethane foam (PUF)/spheroid culture in which the cultured cells spontaneously form spherical multicellular aggregates (spheroids) in the pores of the PUF. We also demonstrate the feasibility of the PUF-HAL system by comparing ES cells to primary hepatocytes in in vitro and ex vivo experiments. Mouse ES cells formed multicellular spheroids in the pores of PUF. ES cells expressed liver-specific functions (ammonia removal and albumin secretion) after treatment with the differentiation-promoting agent, sodium butyrate (SB). We designed a PUF-HAL module comprised of a cylindrical PUF block with many medium-flow capillaries for hepatic differentiation of ES cells. The PUF-HAL module cells expressed ammonia removal and albumin secretion functions after 2 weeks of SB culture. Because of high proliferative activity of ES cells and high cell density, the maximum expression level of albumin secretion function per unit volume of module was comparable to that seen in primary mouse hepatocyte culture. In the animal experiments with rats, the PUF-HAL differentiating ES cells appeared to partially contribute to recovery from liver failure. This outcome indicates that the PUF module containing differentiating ES cells may be a useful biocomponent of a hybrid artificial liver support system.