Development of a hybrid artificial liver using artificial blood

使用人造血液开发混合型人工肝

• 批准号: 10470249
• 负责人: MATSUSHITA Michiaki
• 金额: $ 8.26万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10470249/
• 关键词: Artificial liver; Artificial blood; Two channnel hollow fiber; Porcine; mixed liver cell spheroid; Pyridoxalated-Hemoglobin-Polyoxyethylene conjugate : PHP; 肝細胞スフェロイド; 細胞外基質; コラーゲンゲル; 2系統ホローファイバー; ハイブリッド型人工肝臓; 酸素消費量

This project was aimed to make artificial liver using porcine mixed liver cell sphroids, and to evaluate its function.(1)Mass production of porcine mixed liver cell spheroidWithout chelating agent in cell dispersion, population of non-parenchymal cells were increased, on the contrary, cell viability and yield were decreased. In the collagenase solution, addition of disperse increased the yields and viability of hepatocytes and suppressed the spheroid formation. Production of spheroids could be acheved up to 2.5 x 10^7 cells/ml concentration of hepatocytes in the 1L size spinner flask. Suitable method for porcine liver cell spheroid formation is consisted of rinse with chelating agents, enzyme digestion by collagenase, and spinning culture in 1L sppiner flask with 2.5 x 10^10hepatocytes.(2)Basic experiments : application of artificial blood for artificial liverEvaluation of oxygen transport ability of artificial blood was studied in comparison with red blood cells and culture medium in … More vitro. Oxygen supply capability of artificial blood was equal to that of RBC.In addition, oxygen consumption rate of hepatocyte using artificial blood was the highest of all. Artificial blood could be a way to oxygen supply in artificial liver using hepatocytes.(3)Fabrication and function of artificial liver moduleTwo channnel hollow fiber module, we have already developed, contained 2g amount of spheroids. The one channnel was for oxygenation and the other was for plasma circulation. Pyricoxalated-Hemoglobin-Polyoxyethylene conjugate : PHP was used as an oxygen carrier in the oxygenation circuit and was compared to the culture medium. Anmonium cloride clearance were higher in the PHP group(32% / 2hr)compared to the control group(7.4% / 2hr), and urea and albumin production were also observed. However, PHP easily leaked from the hollow fiber and transit to the other channnel, so the membrane of the fiber must be changed to fiber with smaller pore size. In addition, as the spheroids enclosed in the extra-capillary space aggregated each other, the function of the module was decreased. Therefore, coating or encapsulation of spheroids are considered to be needed for the maintenance of function. Less

本项目旨在利用猪混合肝细胞球体制作人工肝，并对其功能进行评价。(1)猪混合肝细胞球体的批量生产在细胞分散过程中没有使用螯合剂，非实质细胞的数量增加，细胞存活率和产量降低。在胶原酶溶液中，分散体的加入提高了肝细胞的产率和存活率，抑制了球体的形成。在1L大小的转瓶中，肝细胞浓度可达2.5×10~(-7)个/毫升。猪肝细胞球体的适宜形成方法为：用螯合剂冲洗、胶原酶消化、在1L Sppiner瓶中旋转培养2.5×10~(-1)个肝细胞。(2)基础实验：人工血在人工肝中的应用研究…中人工血与红细胞和培养液的氧气输送能力的比较更有试探性。人工血供氧能力与红细胞相当，人工血肝细胞耗氧率最高。人工血可以作为人工肝供氧的一种途径。(3)人工肝模块的研制及功能我们已经研制成功的双通道中空纤维人工肝模块，含有2G的球体。一个通道用于氧合，另一个通道用于血浆循环。吡草酸-血红蛋白-聚氧乙烯偶联物：在氧合回路中使用苯丙氨酸作为氧气载体，并与培养基作比较。与对照组(7.4%/2小时)相比，PHP组(32%/2小时)的氯化铵清除率较高，并观察到尿素和白蛋白的产生。然而，PHP膜很容易从中空纤维中渗漏出来，转移到另一个通道中，因此必须将纤维的膜改为孔径较小的纤维。此外，由于包裹在毛细管外空间中的球体相互聚集，模块的功能降低。因此，球体的涂层或封装被认为是维持功能所必需的。较少

项目成果

齋藤孝則: "早期形成型ラット肝細胞混合スフェロイド機能維持に向けたコラーゲンゲル環境の検討"北海道外科学会誌. 45(1). 90 (2000)

Takanori Saito：“维持早期形成的大鼠肝细胞混合球体功能的胶原凝胶环境的研究”北海道外科学会杂志45(1)。

松下通明: "人工臓器と再生医工学"移植医療,道医シリーズ. 40. 75-78 (2000)

松下道明：“人工器官与再生医学工程”移植医学，土井系列40. 75-78（2000）。

Okubo H.: "Maintenance of morphology and function of mixed liver cell spheroids under collagen gel environment"Journal of Artificial Organs. 4(in press). (2001)

Okubo H.：“胶原凝胶环境下混合肝细胞球体形态和功能的维持”人工器官杂志。

Hisashi Okubo: "Maintenance of morphology and function of mixed liver cell spheroids under collagen gel environment"Journal of Artificial Organs. 4(in press). (2001)

Hisashi Okubo：“胶原凝胶环境下混合肝细胞球体形态和功能的维持”人工器官杂志。

Takanori Saito: "Functional maintenance of mixed liver cell spheroid using collagen gel"The Hokkaido Journal of Surgery. 45(1). 90 (2000)

Takanori Saito：“使用胶原蛋白凝胶维持混合肝细胞球体的功能”北海道外科杂志。