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Development of a new hybrid-type aftificial liver support system using packed-bed type reactor

使用填充床反应器开发新型混合型人工肝支持系统

基本信息

批准号：
05555218
负责人：
OHSHIMA Norio
金额：
$ 6.53万
依托单位：
University of Tsukuba
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

项目摘要

In order to develop a hybrid-type artificial liver support system (ALSS) using cultured hepatocytes, following attempts were carried out.1. Kinetic analyzes of the performance of a hybrid-type ALSSTo obtain sound criteria for the optimal design and improvement of the hybrid-type ALSS using isolated hepatocytes, theoretical and experimental evaluations of the performance of the ALSS were made based on the compartmental mass transfer model and reaction kinetics of the cultured hepatocytes. Results of in vitro and ex vivo perfusion experiments using a hollow-fiber type module, a rotating-disk type module or packed-bed type module indicated that experimentally obtained performance of the module in terms of hepatocyte viability and a few metabolic functions were well correlated with the theoretically estimated performances. Simulation studies related to the design problems of a full-scale device revealed that the performance of the ALSS is inevitably limited by the viability of the hepatocy … More tes. It was suggested that a considerable improvement in the performance of ALSS is to be attained by improving hepatocyte culture conditions.2. Long-term continuous culture of hepatocytes in a packed-bed reactor utilizing porous resinWe investigated the long-term metabolic function of hepatocytes incubated in a newly developed packed-bed type reactor using reticulated polyvinyl formal (PVF) resin as a supporting material. Long-term (up to 1 week) perfusion culture experiments using the packed-bed reactor (20mm i.d.) loaded with 500 PVF resin cubes (mean pore size 250mum, 2*2*2mm), together with conventional monolayr culture experiments as controls, were performed in serum-free or serum-containing medium. Ammonium metabolism, urea synthesis and albumin secretion activities were evaluated quantitatively based on reaction kinetic analyzes as indices of metabolic functions of the cultured hepatocytes. When serum-free medium was used in the perfusion cultures, hepatocytes functions showed significant decay with elapse of the culture period, being less than 10% of those measured on day 1. This loss of activity was more prominent in the perfusion culture than in the monolayr cultures using this medium. In contrast, when serum-containing medium was used, approximately 50% of these activities obtained on day 1 were maintained even at the end of the cultures both in the perfusion and monolayr culture experiments. It was concluded that the packed-bed reactor using PVF resin enabled high-density culture of hepatocytes, and showed a satisfactory ability to maintain metabolic functions of the immobilized hepatocytes for relatively long periods of up to 1 week. This type of reactor is thus considered to represent a breakthrough in overcoming difficulties involved in the development of a hybrid type artificial liver support system. Less

为了开发使用培养的肝细胞的混合型人工肝支持系统（ALSS），进行了以下尝试。混合型人工肝反应器性能的动力学分析为了获得合理的标准，优化设计和改进的混合型人工肝反应器使用离体肝细胞，人工肝反应器的性能进行了理论和实验评估的基础上，房室传质模型和反应动力学培养的肝细胞。在体外和离体灌注实验中使用的中空纤维型模块，一个圆盘型模块或填充床型模块的结果表明，实验获得的性能的模块在肝细胞活力和一些代谢功能的理论估计的性能有很好的相关性。与全尺寸设备的设计问题相关的模拟研究表明，ALSS的性能不可避免地受到肝细胞癌的生存能力的限制。 ...更多信息 tes。提示通过改善肝细胞培养条件，可以显著提高人工肝支持系统的性能.利用多孔树脂在填充床反应器中长期连续培养肝细胞我们研究了以网状聚乙烯醇缩甲醛（PVF）树脂为支撑材料的新型填充床反应器中培养的肝细胞的长期代谢功能。使用填充床反应器（20mm i. d.）加载有500个PVF树脂立方体（平均孔径250 μ m，2 × 2 × 2mm），与作为对照的常规单层培养实验一起，在无血清或含血清培养基中进行。氨代谢，尿素合成和白蛋白分泌活性的定量评价的基础上的反应动力学分析作为指标的代谢功能的培养肝细胞。当在灌注培养中使用无血清培养基时，肝细胞功能随着培养期的推移显示出显著衰减，小于第1天测量的10%。这种活性损失在灌注培养中比在使用该培养基的单层培养中更突出。相反，当使用含血清的培养基时，在灌注和单层培养实验中，即使在培养结束时，在第1天获得的这些活性的约50%也得以维持。得出的结论是，使用PVF树脂的填充床反应器能够高密度培养肝细胞，并显示出令人满意的能力，以维持代谢功能的固定化肝细胞相对较长的时间长达1周。因此，这种类型的反应器被认为代表了克服混合型人工肝支持系统开发中所涉及的困难的突破。少