A preliminary study for constructing a bioartificial liver device with induced pluripotent stem cell-derived hepatocytes.
A preliminary study for constructing a bioartificial liver device with induced pluripotent stem cell-derived hepatocytes.
复制标题
DOI：
10.1186/1475-925x-11-93
复制
发表时间：
2012-12-07
影响因子：
3.9
通讯作者：
Yamamoto K
中科院分区：
文献类型：
--
作者：
Iwamuro M;Shiraha H;Nakaji S;Furutani M;Kobayashi N;Takaki A;Yamamoto K
生物人工肝系统由生物反应器和功能性肝细胞组成，旨在支持肝功能衰竭患者。宿主免疫系统对包埋肝细胞的免疫排斥是一个严重的问题，严重降低了器械的性能。诱导多能干细胞（iPS）被认为是生物人工肝系统的理想来源，因为患者来源的iPS细胞不受免疫排斥。本研究旨在探讨iPS细胞源性肝细胞样细胞构建生物人工肝系统的可行性。小鼠iPS细胞通过多步分化方案经由胚状体和定形内胚层分化为肝细胞样细胞。iPS细胞的分化通过形态学、PCR测定和功能测定来评估。iPS细胞衍生的肝细胞样细胞在孔径为0.2 μm的生物反应器模块中培养7天。通过ELISA分析分泌到循环培养基中的白蛋白的量。此外，在生物反应器模块中培养7天后，通过扫描电子显微镜观察细胞。在分化程序的最后阶段，iPS细胞的形态改变为具有两个核仁和丰富的胞质颗粒的多边形形状。透射电镜分析显示，它们呈多边形，细胞质中有糖原沉积，表面有微绒毛，呈导管状排列。PCR分析显示，在分化过程中白蛋白mRNA的表达增加。还观察到白蛋白和尿素产生。在生物反应器模块中的iPS-Heps培养物显示培养基中白蛋白的积累长达7天。扫描电子显微镜显示附着的细胞簇的中空纤维的模块。这些结果表明，iPS细胞在孔径为0.2 μm的生物反应器模块中培养7天后分化为肝细胞样细胞。我们认为，将具有0.2 μm孔径膜的生物反应器模块与从iPS细胞分化的包埋肝细胞相结合是生物人工肝系统的一个有前途的选择。本文为构建以iPS细胞为导向的生物人工肝系统提供了基本概念和初步数据。PACS代码：87。生物和医学物理，87.85。d生物医学工程，87.85.Lf组织工程，87.85.Tu建模生物医学系统。
Bioartificial liver systems, designed to support patients with liver failure, are composed of bioreactors and functional hepatocytes. Immunological rejection of the embedded hepatocytes by the host immune system is a serious concern that crucially degrades the performance of the device. Induced pluripotent stem (iPS) cells are considered a desirable source for bioartificial liver systems, because patient-derived iPS cells are free from immunological rejection. The purpose of this paper was to test the feasibility of a bioartificial liver system with iPS cell-derived hepatocyte-like cells. Mouse iPS cells were differentiated into hepatocyte-like cells by a multi-step differentiation protocol via embryoid bodies and definitive endoderm. Differentiation of iPS cells was evaluated by morphology, PCR assay, and functional assays. iPS cell-derived hepatocyte-like cells were cultured in a bioreactor module with a pore size of 0.2 μm for 7 days. The amount of albumin secreted into the circulating medium was analyzed by ELISA. Additionally, after a 7-day culture in a bioreactor module, cells were observed by a scanning electron microscope. At the final stage of the differentiation program, iPS cells changed their morphology to a polygonal shape with two nucleoli and enriched cytoplasmic granules. Transmission electron microscope analysis revealed their polygonal shape, glycogen deposition in the cytoplasm, microvilli on their surfaces, and a duct-like arrangement. PCR analysis showed increased expression of albumin mRNA over the course of the differentiation program. Albumin and urea production was also observed. iPS-Heps culture in bioreactor modules showed the accumulation of albumin in the medium for up to 7 days. Scanning electron microscopy revealed the attachment of cell clusters to the hollow fibers of the module. These results indicated that iPS cells were differentiated into hepatocyte-like cells after culture for 7 days in a bioreactor module with a pore size of 0.2 μm. We consider the combination of a bioreactor module with a 0.2-μm pore membrane and embedded hepatocytes differentiated from iPS cells to be a promising option for bioartificial liver systems. This paper provides the basic concept and preliminary data for an iPS cell-oriented bioartificial liver system. PACS code: 87. Biological and medical physics, 87.85.-d Biomedical engineering, 87.85.Lf Tissue engineering, 87.85.Tu Modeling biomedical systems.